indumathi somasundaram | D.Y PATIL UNIVERSITY (original) (raw)

Papers by indumathi somasundaram

Breastfeeding Medicine, 2016

Breastfeeding Medicine, 2016

Human breastmilk is a dynamic, multifaceted biological fluid containing nutrients, bioactive subs... more Human breastmilk is a dynamic, multifaceted biological fluid containing nutrients, bioactive substances, and growth factors. It is effective in supporting growth and development of an infant. As breastmilk has been found to possess mesenchymal stem cells, the importance of the components of breastmilk and their physiological roles is increasing day by day. The present study was intended to identify the secretions of growth factors, mainly vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF), from human breastmilk mesenchymal stem cells under basal conditions of in vitro cell culture using synthetic media and human cord serum. The growth factors were analyzed with the enzyme-linked immunosorbent assay technique. The cultured mesenchymal stem cells of breastmilk without serum revealed significant differences in secretions of the VEGF and HGF growth factors (8.55 ± 2.26402 pg/mL and 230.8 ± 45.9861 pg/mL, respectively) compared with mesenchymal stem cells of breastmilk with serum (21.31 ± 4.69 pg/mL and 2,404.42 ± 481.593 pg/mL, respectively). Results obtained from our study demonstrate that both VEGF and HGF are secreted in vitro by human breastmilk mesenchymal stem cells. The roles of VEGF and HGF in surfactant secretion, pulmonary maturation, and neonatal maturity have been well established. Thus, we emphasize that breastmilk-derived MSCs could be a potent therapeutic source in treating neonatal diseases. Besides, due to its immense potency, the study also emphasizes the importance of breastfeeding, which is promoted by organizations like the World Heatlh Organization and UNICEF.

The widely accepted source of stem cell for clinical transplant is the infusion of mononuclear ce... more The widely accepted source of stem cell for clinical transplant is the infusion of mononuclear cell (MNC) from bone marrow. The disadvantages of MNC infusion pose a threat to the world of regenerative medicine because of its vast heterogeneity, granulocyte interface and age. Thus, finding an alternative on improvisation of MNC therapy is of utmost importance. Thereby, we hypothesized an alternative approach by sorting of lymphocytes and monocytes using an FACS based purification method. To confirm our speculation, we enumerated various stem cell populations in non-sorted as well as sorted MNC from bone marrow. We found that sorted lymphocytes and monocytes possess high percentage of stem cells when compared to non-sorted MNC. Thus, it is concluded that sorted lymphocytes and monocytes cocktail from bone marrow might be an efficient alternative approach in cell based therapies. However, further investigations on these sorted cells will bring this work closer to clinical applications....

Research on adult stem cells has been a great deal of excitement. The candidate stem cells presen... more Research on adult stem cells has been a great deal of excitement. The candidate stem cells present in the adult tissues are the hematopoietic and mesenchymal stem cells. However, there exist other heterogenous cell such as side population (SP) and endothelial progenitors (EP) that has the properties of repair and regeneration. Much work pertaining to these subpopulations is at its infancy and more research in clinical practice is of utmost important. Thus, the objective of this work is to find out whether subpopulations exist in bone marrow in higher percentage and play a vital role in tissue repair and regeneration. To test this hypothesis, we characterized stem cell populations versus subpopulations including EP cells and SP cells from Bone Marrow samples (n = 5). Surprisingly, we found that the mean of Endoglin CD105+ CD34-, CD105+ CD90 - for EP cells was found to be 41.68% and 35.88% respectively. The mean of ABCG2+ and CD117+ SP cells was found to be 3.58% and 4.34%. These resu...

There has been nearly a century long search for the elusive cure of diabetes mellitus. While insu... more There has been nearly a century long search for the elusive cure of diabetes mellitus. While insulin replacement remains the cornerstone treatment for diabetes mellitus, the transplantation of pancreatic islets of Langerhans has the potential to become an important alternative. However, islet transplantation has been hampered by a shortage of donors and immune rejection. Stem cells have been an enigmatic area of research and progressive effort has been made in order to provide treatment for various devastating disorders, inclusive of diabetes mellitus. Among the promising sources of autologous stem cells, major strides have been made to generate insulin producing cells in vitro from bone marrow and adipose tissue. Several exciting reports had emerged exploring the potential of these aforesaid stem cells as an alternative diabetic therapy. Despite continuous efforts and rigorous assessments, there has been scepticism in accepting stem cells as therapy for diabetes. We herein disclose...

El Mednifico Journal, 2014

Stem Cell Therapy for Organ Failure, 2014

Stem Cell Therapy for Organ Failure, 2014

Stem Cell Therapy for Organ Failure, 2014

Stem Cells International, 2015

The study aims to identify the phenotypic marker expressions of different human adult stem cells ... more The study aims to identify the phenotypic marker expressions of different human adult stem cells derived from, namely, bone marrow, subcutaneous fat, and omentum fat, cultured in different media, namely, DMEM-Low Glucose, Alpha-MEM, DMEM-F12 and DMEM-KO and under long term culture conditions (>P20). We characterized immunophenotype by using various hematopoietic, mesenchymal, endothelial markers, and cell adhesion molecules in the long term cultures (Passages-P1, P3, P5, P9, P12, P15, and P20.) Interestingly, data revealed similar marker expression profiles irrespective of source, basal media, and extensive culturing. This demonstrates that all adult stem cell sources mentioned in this study share similar phenotypic marker and all media seem appropriate for culturing these sources. However, a disparity was observed in the markers such as CD49d, CD54, CD117, CD29, and CD106, thereby warranting further research on these markers. Besides the aforesaid objective, it is understood from the study that immunophenotyping acts as a valuable tool to identify inherent property of each cell, thereby leading to a valuable cell based therapy.

Stem Cells in Aesthetic Procedures, 2014

International Journal of Biology, 2010

Adipose tissue represents an abundant and accessible source of multipotent adult stem cells, whic... more Adipose tissue represents an abundant and accessible source of multipotent adult stem cells, which appear to possess a yet-undetermined degree of plasticity. Subcutaneous adipose tissue is much studied in recent years than omentum fat tissue. Hence the objective of this work is to characterize the freshly isolated human adipose tissue derived stromal vascular fraction cell relative to passage 0 from subcutaneous fat and the omentum fat. The stromal vascular fraction of both subcutaneous and omentum fat contains hematopoietic and mesenchymal stem cell population where as in P0 the mesenchymal stem cell alone was retained. The side population ABCG2 is present in subcutaneous fat, whereas no ABCG2 expression is found in omentum fat. This study showed that the omentum fat exhibit higher percentage of hematopoietic cells compared to subcutaneous fat in both SVF and P0. It is concluded from this study that not only subcutaneous fat, but also omentum fat derived stem cells serve as a therapeutic potential in Regenerative medicine.

Tissue and Cell, 2013

Scientific explorations on feto-maternal organ stem cells revealed its possible applicability in ... more Scientific explorations on feto-maternal organ stem cells revealed its possible applicability in treatment of various diseases. However, establishment of an ideal placental tissue stem cell source in regenerative application is inconclusive and arduous. Hence, this study aims to resolve this tribulation by comparison of mesenchymal stem cells (MSC) from fetal placenta - amniotic membrane (AM-MSC), chorionic plate (CP-MSC) tissue and the maternal placenta-Decidua (D-MSC), thereby facilitating the researchers to determine their pertinent source. The cells were expanded and scrutinized for expression profiling, proliferation and differentiation ability. Remarkable expressions of certain markers in addition to its prospective mesodermal differentiation confirmed their mesenchyme origin. Despite the specified alikeness among these sources, reliable and non-invasive procurement of AM-MSC coupled with its higher growth potency makes it the most constructive stem cell source. However, exhibited similarities demands further investigations on extensive expandability and cytogenetic stability of these sources prior to its therapeutic applicability.

Cytotechnology, 2013

Bone marrow derived stem cells (BMSC) have paved way to clinical approaches for its utilization i... more Bone marrow derived stem cells (BMSC) have paved way to clinical approaches for its utilization in a variety of diseases due to its ease of isolation combined with its multilineage differentiation capacity. However, the applicability of BMSC is not successful due to the lesser number of nucleated cells obtained from large samples. Hence, culture expansion of BMSC is a prerequisite, as high numbers of stem cells are needed to meet the standards of clinical advancement. There are attempts on optimizing culture condition for large scale production of BMSC. It was believed that, prolonged culture of BMSC is difficult since they tend to lose their characteristics and differentiation potential. Hence, our study aims to determine whether BMSCs could retain its proliferative and differentiation capacity in prolonged in vitro culture by a comparative study on extensive culturing of BMSC with the following four media, DMEM LG (DMEM-Low Glucose), DMEM KO (DMEM-Knock Out), Alpha MEM (Alpha Minimal Essential Medium), DMEM F 12. We found that two samples among the three cultured tend to lose their property in long term culturing. Besides, we also found that DMEM LG and Alpha MEM were the optimal media for in vitro culturing of BMSC. Overall, it was concluded that BMSC can be cultured until passage 15 without losing its characteristics. However, its potency beyond passage 15 has to be further elucidated for utilization of the ex vivo expanded BMSC for subsequent cellular therapies.

Cytotechnology, 2013

The immense potency of nutritional components of human breast milk and importance of breastfeedin... more The immense potency of nutritional components of human breast milk and importance of breastfeeding is known worldwide. Recent researches had identified stem cells as integral component of human breast milk. Nevertheless, there is little proof of evidence on the stem cell constituents of breast milk. It is imperative to explore the cellular constituents of human breast milk, including of stem cells, to open new avenue in child's development and regeneration. Thus, we aimed at identifying the cellular constituents of human breast milk by phenotypic characterisation of diverse cell surface markers of hematopoietic stem cells (CD 34, CD 133, CD 117), mesenchymal stem cells (CD 90, CD 105, CD 73), myoepithelial cells (CD 29, CD 44), Immune cells (CD 209, CD 86, CD 83, CD 14, CD 13, HLADR, CD 45), as well as cell adhesion molecules (CD 31, CD 54, CD 166, CD 106, CD 49d), and other markers (ABCG2, CD140b) using flowcytometry. We found a lower expression of CD 34 (13.07 ± 2.0 %), CD 90 (7.79 ± 0.8 %) and CD 73 (2.19 ± 0.41 %), indicating scanty hematopoietic and mesenchymal stem cell population in human breast milk. On contrary, myoepithelial progenitors, cell adhesion molecules, immune cells and growth factors were identified as the major constituents of breast milk. Overall, this study illuminates the benefits of breast feeding as breast milk encompasses heterogeneous cellular components that benefits child's growth, immunity and development. However, further research on these constituents of human breast milk will widen their applicability in treatment of neonatal disorders.

Cytotechnology, 2012

Omentum fat derived stem cells have emerged as an alternative and accessible therapeutic tool in ... more Omentum fat derived stem cells have emerged as an alternative and accessible therapeutic tool in recent years in contrast to the existing persuasive sources of stem cells, bone marrow and subcutaneous adipose tissue. However, there has been a scanty citation on human omentum fat derived stem cells. Furthermore, identification of specific cell surface markers among aforesaid sources is still controversial. In lieu of this existing perplexity, the current research work aims at signifying omentum fat as a ground-breaking source of stem cells by surface antigenic profiling of stem cell population. In this study, we examined and compared the profiling of cell surface antigenic expressions of hematopoietic stem cells, mesenchymal stem cells, cell adhesion molecules and other unique markers such as ABCG2, ALDH and CD 117 in whole cell population of human omentum fat, subcutaneous fat and bone marrow. The phenotypic characterization through flowcytometry revealed the positive expressions of CD 34, CD 45, CD 133, HLADR, CD 90, CD 105, CD 73, CD 29, CD 13, CD 44, CD 54, CD 31, ALDH and CD 117 in all sources. The similarities between the phenotypic expressions of omentum fat derived stem cells to that of subcutaneous fat and bone marrow substantiates that identification of ultimate source for curative therapeutics is arduous to assess. Nevertheless, these results support the potential therapeutic application of omentum fat derived stem cells.

Cytotechnology, 2013

Human umbilical cord blood (hUCB) has been the preferred source of stem cells for the treatment o... more Human umbilical cord blood (hUCB) has been the preferred source of stem cells for the treatment of haematological malignancies and genetic disorders. This is primarily due to its non-invasiveness, high accessibility with relative ease of isolation. Still failures do prevail due to its heterogeneity and lesser frequency of MSC identified in UCB. This study, thus, employs a cell enrichment technology to improve its therapeutic efficacy. This was achieved by immunophenotypic comparison of stem cells isolated from the heterogenous non-sorted mononuclear cells (MNCs), linage depleted (Lin+ and Lin-) fractions obtained from magnetic activated cell sorter (MACS) and sorted MNCs obtained by fluorescent activated cell sorter (FACS). The markers under consideration were CD29, CD44, CD34, CD45, CD133, CD90 and CD117. FACS sorted MNCs were rich in naive stem cell population, whereas non-sorted MNCs and lineage depleted fractions were found to be rich in progenitors. Thus, we suggest that a combination therapy of both sorted population might serve as an alternative valuable tool in treating haematologic/genetic disorders. However, further research on cell enrichment technology might give a clue for improved cell based therapy in regenerative medicine.

Cytotechnology, 2014

The therapeutic rationale for tissue repair and regeneration using stem cells is at its infancy a... more The therapeutic rationale for tissue repair and regeneration using stem cells is at its infancy and needs advancement in understanding the role of individual component's innate capability. As stem cells of adipose tissue reside in a more heterogeneous population of stromal vascular fractions, cell separation or sorting becomes an eminent step towards revealing their unique properties. This study elucidates the comparative efficacy of lineage depleted adipose derived stromal vascular fraction (SVF) and their innate ability using magnetic activated cell sorter (MACS). To this end, isolated SVF from human adipose tissue was lineage depleted according to the manufacturer's instructions using specific antibody cocktail through MACS. The enriched lineage negative (lin-) and lineage positive (lin+) cell fractions were cultured, phenotypically characterized for the panel of cell surface markers using flowcytometry and subjected to osteoblastic and adipogenic differentiation. The expression profile obtained for lin- cells was CD34-/CD45-/HLADR-/CD49d-/CD140b-/CD31-/CD90+/CD105+/CD73+/CD54+/CD166+/CD117- when compared to Lin+ cells expressing CD34+/CD45+/HLADR-/CD49d-/CD140b+/CD31-/CD90+/CD105+/CD73+/CD54+/CD166+/CD117+ (CD-cluster of differentiation). These results, thus, advances our understanding on the inherent property of the individual cell population. Furthermore, both the fractions exhibited mesodermal lineage differentiation capacity. To conclude, this research pursuit rationalized the regenerative therapeutic applicability of both lin- and lin+ cultures of human adipose tissue for disorders of mesodermal, haematological and vascular origin.

Cytokine, 2012

The process of the growth of the fetus begins in the uterus and gets further accelerated followin... more The process of the growth of the fetus begins in the uterus and gets further accelerated following the birth, especially during initial few months. The role of the growth factors in the physiology of the cellular growth is already well established. Vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) seem to be imperative for angiogenesis, cell development and proliferation as well as maintenance of the tissues. The levels of these factors in the maternal serum during pregnancy as well as during postpartum period are insignificant. Consequently, we hypothesized that the fetus receives moderate supply of these growth factors from the placenta during its stay in the uterus. This supply gets further augmented during the postpartum period through the different source, i.e. mother's milk. To study this physiological transition of the source of the growth factors from the placenta to the breast milk, the concentrations of VEGF and HGF in the cord serum of full term neonates and that in the breast milk of the corresponding mothers were analyzed during ELISA. The human milk, especially the colostrum revealed significantly higher levels of VEGF and HGF (1541.759 ± 119.349 pg/ml and 7129.249 ± 273.472 pg/ml) than cord serum (16.632 ± 0.773 pg/ml and 2581.6 ± 108.275 pg/ml) respectively. The multifold higher levels of VEGF observed in colostrum probably correlates with its high neonatal requirement for the maturation of the gastrointestinal epithelium following birth. The higher levels of both the growth factors in the breast milk than those observed in the cord serum probably explain their higher needs by the neonates for immunological protection, protein synthesis and neurocognitive development. The observations of the present study strengthen the policy of the colostrum feeding, which is promoted by organizations like World Health Organization (WHO). This study further documents the fact that the commercial milk formulae cannot replace the human milk.

Cellular & Molecular Biology Letters, 2013

Breastfeeding Medicine, 2016

Breastfeeding Medicine, 2016

Human breastmilk is a dynamic, multifaceted biological fluid containing nutrients, bioactive subs... more Human breastmilk is a dynamic, multifaceted biological fluid containing nutrients, bioactive substances, and growth factors. It is effective in supporting growth and development of an infant. As breastmilk has been found to possess mesenchymal stem cells, the importance of the components of breastmilk and their physiological roles is increasing day by day. The present study was intended to identify the secretions of growth factors, mainly vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF), from human breastmilk mesenchymal stem cells under basal conditions of in vitro cell culture using synthetic media and human cord serum. The growth factors were analyzed with the enzyme-linked immunosorbent assay technique. The cultured mesenchymal stem cells of breastmilk without serum revealed significant differences in secretions of the VEGF and HGF growth factors (8.55 ± 2.26402 pg/mL and 230.8 ± 45.9861 pg/mL, respectively) compared with mesenchymal stem cells of breastmilk with serum (21.31 ± 4.69 pg/mL and 2,404.42 ± 481.593 pg/mL, respectively). Results obtained from our study demonstrate that both VEGF and HGF are secreted in vitro by human breastmilk mesenchymal stem cells. The roles of VEGF and HGF in surfactant secretion, pulmonary maturation, and neonatal maturity have been well established. Thus, we emphasize that breastmilk-derived MSCs could be a potent therapeutic source in treating neonatal diseases. Besides, due to its immense potency, the study also emphasizes the importance of breastfeeding, which is promoted by organizations like the World Heatlh Organization and UNICEF.

The widely accepted source of stem cell for clinical transplant is the infusion of mononuclear ce... more The widely accepted source of stem cell for clinical transplant is the infusion of mononuclear cell (MNC) from bone marrow. The disadvantages of MNC infusion pose a threat to the world of regenerative medicine because of its vast heterogeneity, granulocyte interface and age. Thus, finding an alternative on improvisation of MNC therapy is of utmost importance. Thereby, we hypothesized an alternative approach by sorting of lymphocytes and monocytes using an FACS based purification method. To confirm our speculation, we enumerated various stem cell populations in non-sorted as well as sorted MNC from bone marrow. We found that sorted lymphocytes and monocytes possess high percentage of stem cells when compared to non-sorted MNC. Thus, it is concluded that sorted lymphocytes and monocytes cocktail from bone marrow might be an efficient alternative approach in cell based therapies. However, further investigations on these sorted cells will bring this work closer to clinical applications....

Research on adult stem cells has been a great deal of excitement. The candidate stem cells presen... more Research on adult stem cells has been a great deal of excitement. The candidate stem cells present in the adult tissues are the hematopoietic and mesenchymal stem cells. However, there exist other heterogenous cell such as side population (SP) and endothelial progenitors (EP) that has the properties of repair and regeneration. Much work pertaining to these subpopulations is at its infancy and more research in clinical practice is of utmost important. Thus, the objective of this work is to find out whether subpopulations exist in bone marrow in higher percentage and play a vital role in tissue repair and regeneration. To test this hypothesis, we characterized stem cell populations versus subpopulations including EP cells and SP cells from Bone Marrow samples (n = 5). Surprisingly, we found that the mean of Endoglin CD105+ CD34-, CD105+ CD90 - for EP cells was found to be 41.68% and 35.88% respectively. The mean of ABCG2+ and CD117+ SP cells was found to be 3.58% and 4.34%. These resu...

There has been nearly a century long search for the elusive cure of diabetes mellitus. While insu... more There has been nearly a century long search for the elusive cure of diabetes mellitus. While insulin replacement remains the cornerstone treatment for diabetes mellitus, the transplantation of pancreatic islets of Langerhans has the potential to become an important alternative. However, islet transplantation has been hampered by a shortage of donors and immune rejection. Stem cells have been an enigmatic area of research and progressive effort has been made in order to provide treatment for various devastating disorders, inclusive of diabetes mellitus. Among the promising sources of autologous stem cells, major strides have been made to generate insulin producing cells in vitro from bone marrow and adipose tissue. Several exciting reports had emerged exploring the potential of these aforesaid stem cells as an alternative diabetic therapy. Despite continuous efforts and rigorous assessments, there has been scepticism in accepting stem cells as therapy for diabetes. We herein disclose...

El Mednifico Journal, 2014

Stem Cell Therapy for Organ Failure, 2014

Stem Cell Therapy for Organ Failure, 2014

Stem Cell Therapy for Organ Failure, 2014

Stem Cells International, 2015

The study aims to identify the phenotypic marker expressions of different human adult stem cells ... more The study aims to identify the phenotypic marker expressions of different human adult stem cells derived from, namely, bone marrow, subcutaneous fat, and omentum fat, cultured in different media, namely, DMEM-Low Glucose, Alpha-MEM, DMEM-F12 and DMEM-KO and under long term culture conditions (>P20). We characterized immunophenotype by using various hematopoietic, mesenchymal, endothelial markers, and cell adhesion molecules in the long term cultures (Passages-P1, P3, P5, P9, P12, P15, and P20.) Interestingly, data revealed similar marker expression profiles irrespective of source, basal media, and extensive culturing. This demonstrates that all adult stem cell sources mentioned in this study share similar phenotypic marker and all media seem appropriate for culturing these sources. However, a disparity was observed in the markers such as CD49d, CD54, CD117, CD29, and CD106, thereby warranting further research on these markers. Besides the aforesaid objective, it is understood from the study that immunophenotyping acts as a valuable tool to identify inherent property of each cell, thereby leading to a valuable cell based therapy.

Stem Cells in Aesthetic Procedures, 2014

International Journal of Biology, 2010

Adipose tissue represents an abundant and accessible source of multipotent adult stem cells, whic... more Adipose tissue represents an abundant and accessible source of multipotent adult stem cells, which appear to possess a yet-undetermined degree of plasticity. Subcutaneous adipose tissue is much studied in recent years than omentum fat tissue. Hence the objective of this work is to characterize the freshly isolated human adipose tissue derived stromal vascular fraction cell relative to passage 0 from subcutaneous fat and the omentum fat. The stromal vascular fraction of both subcutaneous and omentum fat contains hematopoietic and mesenchymal stem cell population where as in P0 the mesenchymal stem cell alone was retained. The side population ABCG2 is present in subcutaneous fat, whereas no ABCG2 expression is found in omentum fat. This study showed that the omentum fat exhibit higher percentage of hematopoietic cells compared to subcutaneous fat in both SVF and P0. It is concluded from this study that not only subcutaneous fat, but also omentum fat derived stem cells serve as a therapeutic potential in Regenerative medicine.

Tissue and Cell, 2013

Scientific explorations on feto-maternal organ stem cells revealed its possible applicability in ... more Scientific explorations on feto-maternal organ stem cells revealed its possible applicability in treatment of various diseases. However, establishment of an ideal placental tissue stem cell source in regenerative application is inconclusive and arduous. Hence, this study aims to resolve this tribulation by comparison of mesenchymal stem cells (MSC) from fetal placenta - amniotic membrane (AM-MSC), chorionic plate (CP-MSC) tissue and the maternal placenta-Decidua (D-MSC), thereby facilitating the researchers to determine their pertinent source. The cells were expanded and scrutinized for expression profiling, proliferation and differentiation ability. Remarkable expressions of certain markers in addition to its prospective mesodermal differentiation confirmed their mesenchyme origin. Despite the specified alikeness among these sources, reliable and non-invasive procurement of AM-MSC coupled with its higher growth potency makes it the most constructive stem cell source. However, exhibited similarities demands further investigations on extensive expandability and cytogenetic stability of these sources prior to its therapeutic applicability.

Cytotechnology, 2013

Bone marrow derived stem cells (BMSC) have paved way to clinical approaches for its utilization i... more Bone marrow derived stem cells (BMSC) have paved way to clinical approaches for its utilization in a variety of diseases due to its ease of isolation combined with its multilineage differentiation capacity. However, the applicability of BMSC is not successful due to the lesser number of nucleated cells obtained from large samples. Hence, culture expansion of BMSC is a prerequisite, as high numbers of stem cells are needed to meet the standards of clinical advancement. There are attempts on optimizing culture condition for large scale production of BMSC. It was believed that, prolonged culture of BMSC is difficult since they tend to lose their characteristics and differentiation potential. Hence, our study aims to determine whether BMSCs could retain its proliferative and differentiation capacity in prolonged in vitro culture by a comparative study on extensive culturing of BMSC with the following four media, DMEM LG (DMEM-Low Glucose), DMEM KO (DMEM-Knock Out), Alpha MEM (Alpha Minimal Essential Medium), DMEM F 12. We found that two samples among the three cultured tend to lose their property in long term culturing. Besides, we also found that DMEM LG and Alpha MEM were the optimal media for in vitro culturing of BMSC. Overall, it was concluded that BMSC can be cultured until passage 15 without losing its characteristics. However, its potency beyond passage 15 has to be further elucidated for utilization of the ex vivo expanded BMSC for subsequent cellular therapies.

Cytotechnology, 2013

The immense potency of nutritional components of human breast milk and importance of breastfeedin... more The immense potency of nutritional components of human breast milk and importance of breastfeeding is known worldwide. Recent researches had identified stem cells as integral component of human breast milk. Nevertheless, there is little proof of evidence on the stem cell constituents of breast milk. It is imperative to explore the cellular constituents of human breast milk, including of stem cells, to open new avenue in child's development and regeneration. Thus, we aimed at identifying the cellular constituents of human breast milk by phenotypic characterisation of diverse cell surface markers of hematopoietic stem cells (CD 34, CD 133, CD 117), mesenchymal stem cells (CD 90, CD 105, CD 73), myoepithelial cells (CD 29, CD 44), Immune cells (CD 209, CD 86, CD 83, CD 14, CD 13, HLADR, CD 45), as well as cell adhesion molecules (CD 31, CD 54, CD 166, CD 106, CD 49d), and other markers (ABCG2, CD140b) using flowcytometry. We found a lower expression of CD 34 (13.07 ± 2.0 %), CD 90 (7.79 ± 0.8 %) and CD 73 (2.19 ± 0.41 %), indicating scanty hematopoietic and mesenchymal stem cell population in human breast milk. On contrary, myoepithelial progenitors, cell adhesion molecules, immune cells and growth factors were identified as the major constituents of breast milk. Overall, this study illuminates the benefits of breast feeding as breast milk encompasses heterogeneous cellular components that benefits child's growth, immunity and development. However, further research on these constituents of human breast milk will widen their applicability in treatment of neonatal disorders.

Cytotechnology, 2012

Omentum fat derived stem cells have emerged as an alternative and accessible therapeutic tool in ... more Omentum fat derived stem cells have emerged as an alternative and accessible therapeutic tool in recent years in contrast to the existing persuasive sources of stem cells, bone marrow and subcutaneous adipose tissue. However, there has been a scanty citation on human omentum fat derived stem cells. Furthermore, identification of specific cell surface markers among aforesaid sources is still controversial. In lieu of this existing perplexity, the current research work aims at signifying omentum fat as a ground-breaking source of stem cells by surface antigenic profiling of stem cell population. In this study, we examined and compared the profiling of cell surface antigenic expressions of hematopoietic stem cells, mesenchymal stem cells, cell adhesion molecules and other unique markers such as ABCG2, ALDH and CD 117 in whole cell population of human omentum fat, subcutaneous fat and bone marrow. The phenotypic characterization through flowcytometry revealed the positive expressions of CD 34, CD 45, CD 133, HLADR, CD 90, CD 105, CD 73, CD 29, CD 13, CD 44, CD 54, CD 31, ALDH and CD 117 in all sources. The similarities between the phenotypic expressions of omentum fat derived stem cells to that of subcutaneous fat and bone marrow substantiates that identification of ultimate source for curative therapeutics is arduous to assess. Nevertheless, these results support the potential therapeutic application of omentum fat derived stem cells.

Cytotechnology, 2013

Human umbilical cord blood (hUCB) has been the preferred source of stem cells for the treatment o... more Human umbilical cord blood (hUCB) has been the preferred source of stem cells for the treatment of haematological malignancies and genetic disorders. This is primarily due to its non-invasiveness, high accessibility with relative ease of isolation. Still failures do prevail due to its heterogeneity and lesser frequency of MSC identified in UCB. This study, thus, employs a cell enrichment technology to improve its therapeutic efficacy. This was achieved by immunophenotypic comparison of stem cells isolated from the heterogenous non-sorted mononuclear cells (MNCs), linage depleted (Lin+ and Lin-) fractions obtained from magnetic activated cell sorter (MACS) and sorted MNCs obtained by fluorescent activated cell sorter (FACS). The markers under consideration were CD29, CD44, CD34, CD45, CD133, CD90 and CD117. FACS sorted MNCs were rich in naive stem cell population, whereas non-sorted MNCs and lineage depleted fractions were found to be rich in progenitors. Thus, we suggest that a combination therapy of both sorted population might serve as an alternative valuable tool in treating haematologic/genetic disorders. However, further research on cell enrichment technology might give a clue for improved cell based therapy in regenerative medicine.

Cytotechnology, 2014

The therapeutic rationale for tissue repair and regeneration using stem cells is at its infancy a... more The therapeutic rationale for tissue repair and regeneration using stem cells is at its infancy and needs advancement in understanding the role of individual component's innate capability. As stem cells of adipose tissue reside in a more heterogeneous population of stromal vascular fractions, cell separation or sorting becomes an eminent step towards revealing their unique properties. This study elucidates the comparative efficacy of lineage depleted adipose derived stromal vascular fraction (SVF) and their innate ability using magnetic activated cell sorter (MACS). To this end, isolated SVF from human adipose tissue was lineage depleted according to the manufacturer's instructions using specific antibody cocktail through MACS. The enriched lineage negative (lin-) and lineage positive (lin+) cell fractions were cultured, phenotypically characterized for the panel of cell surface markers using flowcytometry and subjected to osteoblastic and adipogenic differentiation. The expression profile obtained for lin- cells was CD34-/CD45-/HLADR-/CD49d-/CD140b-/CD31-/CD90+/CD105+/CD73+/CD54+/CD166+/CD117- when compared to Lin+ cells expressing CD34+/CD45+/HLADR-/CD49d-/CD140b+/CD31-/CD90+/CD105+/CD73+/CD54+/CD166+/CD117+ (CD-cluster of differentiation). These results, thus, advances our understanding on the inherent property of the individual cell population. Furthermore, both the fractions exhibited mesodermal lineage differentiation capacity. To conclude, this research pursuit rationalized the regenerative therapeutic applicability of both lin- and lin+ cultures of human adipose tissue for disorders of mesodermal, haematological and vascular origin.

Cytokine, 2012

The process of the growth of the fetus begins in the uterus and gets further accelerated followin... more The process of the growth of the fetus begins in the uterus and gets further accelerated following the birth, especially during initial few months. The role of the growth factors in the physiology of the cellular growth is already well established. Vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) seem to be imperative for angiogenesis, cell development and proliferation as well as maintenance of the tissues. The levels of these factors in the maternal serum during pregnancy as well as during postpartum period are insignificant. Consequently, we hypothesized that the fetus receives moderate supply of these growth factors from the placenta during its stay in the uterus. This supply gets further augmented during the postpartum period through the different source, i.e. mother's milk. To study this physiological transition of the source of the growth factors from the placenta to the breast milk, the concentrations of VEGF and HGF in the cord serum of full term neonates and that in the breast milk of the corresponding mothers were analyzed during ELISA. The human milk, especially the colostrum revealed significantly higher levels of VEGF and HGF (1541.759 ± 119.349 pg/ml and 7129.249 ± 273.472 pg/ml) than cord serum (16.632 ± 0.773 pg/ml and 2581.6 ± 108.275 pg/ml) respectively. The multifold higher levels of VEGF observed in colostrum probably correlates with its high neonatal requirement for the maturation of the gastrointestinal epithelium following birth. The higher levels of both the growth factors in the breast milk than those observed in the cord serum probably explain their higher needs by the neonates for immunological protection, protein synthesis and neurocognitive development. The observations of the present study strengthen the policy of the colostrum feeding, which is promoted by organizations like World Health Organization (WHO). This study further documents the fact that the commercial milk formulae cannot replace the human milk.

Cellular & Molecular Biology Letters, 2013