Characterization of Human Adipose Tissue Derived Hematopoietic Stem Cell, Mesenchymal Stem Cell and Side Population Cells (original) (raw)
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Cytotherapy, 2013
Background aims. Adipose tissue is a rich and very convenient source of cells for regenerative medicine therapeutic approaches. However, a characterization of the population of adipose-derived stromal and stem cells (ASCs) with the greatest therapeutic potential remains unclear. Under the authority of International Federation of Adipose Therapeutics and International Society for Cellular Therapy, this paper sets out to establish minimal definitions of stromal cells both as uncultured stromal vascular fraction (SVF) and as an adherent stromal/stem cells population. Methods. Phenotypic and functional criteria for the identification of adipose-derived cells were drawn from the literature. Results. In the SVF, cells are identified phenotypically by the following markers: CD45-CD235a-CD31-CD34þ. Added value may be provided by both a viability marker and the following surface antigens: CD13, CD73, CD90 and CD105. The fibroblastoid colony-forming unit assay permits the evaluation of progenitor frequency in the SVF population. In culture, ASCs retain markers in common with other mesenchymal stromal/stem cells (MSCs), including CD90, CD73, CD105, and CD44 and remain negative for CD45 and CD31. They can be distinguished from bone-marrow-derived MSCs by their positivity for CD36 and negativity for CD106. The CFU-F assay is recommended to calculate population doublings capacity of ASCs. The adipocytic, chondroblastic and osteoblastic differentiation assays serve to complete the cell identification and potency assessment in conjunction with a quantitative evaluation of the differentiation either biochemically or by reverse transcription polymerase chain reaction. Conclusions. The goal of this paper is to provide initial guidance for the scientific community working with adipose-derived cells and to facilitate development of international standards based on reproducible parameters.
The FASEB Journal, 2009
Adipose tissue is a dynamic endocrine organ with a central role in metabolism regulation. Functional differences in adipose tissue seem associated with the regional distribution of fat depots, in particular in subcutaneous and visceral omental pads. Here, we report for the first time the isolation of human adipose-derived adult stem cells from visceral omental and subcutaneous fat (V-ASCs and S-ASCs, respectively) from the same subject. Immunophenotyping shows that plastic culturing selects homogeneous cell populations of V-ASCs and S-ASCs from the corresponding stromal vascular fractions (SVFs), sharing typical markers of mesenchymal stem cells. Electron microscopy and electrophysiological and real-time RT-PCR analyses confirm the mesenchymal stem nature of both V-ASCs and S-ASCs, while no significant differences in a limited pattern of cytokine/chemokine expression can be detected. Similar to S-ASCs, V-ASCs can differentiate in vitro toward adipogenic, osteogenic, chondrogenic, muscular, and neuronal lineages, as demonstrated by histochemical, immunofluorescence, real-time RT-PCR, and electrophysiological analyses, suggesting the multipotency of such adult stem cells. Our data demonstrate that both visceral and subcutaneous adipose tissues are a source of pluripotent stem cells with multigermline potential. However, the visceral rather than the subcutaneous ASC could represent a more appropriate in vitro cell model for investigating the molecular mechanisms implicated in the pathophysiology of metabolic disorders such as obesityterization of human adult stem-cell populations isolated from visceral and subcutaneous adipose tissue. FASEB J. 23, 000 -000 (2009). www.fasebj.org 1 0892-6638/09/0023-0001 © FASEB The FASEB Journal article fj.08-126946. Published online July 7, 2009.
Aesthetic Plastic Surgery, 2020
Background New regenerative treatments have emerged with the use of multipotent mesenchymal cells, with special interest in adipose-derived stem cells (ADSCs). In recent years, studies that have sought to identify possible quantitative or qualitative differences in ADSCs derived from different donor subcutaneous adipose tissue have shown divergent results making the determination of a preferential donor area still considered inconclusive. Materials and Methods The number of ADSCs present in the adipose tissue collected by liposuction was quantified between five different body areas from 17 women, by means of the CFU-F assay and to investigate possible qualitative differences in the ADSCs from these different areas by analyzing: cell surface markers, cell kinetics, action of the supernatant produced by ADSCs from different body areas on fibroblast migration and, finally, differences in the secretome present in the supernatant produced by these cells. Results The highest mean concentration of CFU-Fs was the dorsum (23.20 ± 26.13), and the lowest was the thighs (6.87 ± 5.04). No qualitative differences were observed in the expression of the cell surface markers or in cell kinetics. Supernatants produced by the ADSCs derived from the abdomen and the thighs demonstrated an increased rate of migration of fibroblasts in vitro similarly. No differences were observed in the secretome between the ADSCs groups. Conclusions It was observed that the region of the dorsal upper back presented a greater number of ADSCs than the thighs. No qualitative differences were observed between the ADSCs of the five areas analyzed. No Level Assigned This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266\. Keywords Adipose-derived stem cells Á Stem cells Á Adipose tissue Á Subcutaneous adipose tissue body donor areas Á Body donor areas Á Fibroblastic migration Á Fibroblasts Á Adipose derived stem cell secretome
Cytotherapy, 2010
Background aims . Adipose-derived stromal/stem cells (ASC) capable of multipotential differentiation can be isolated with high yields from human subcutaneous lipoaspirates. This study reports our recent experience of isolating and immunophenotypically characterizing ASC from Ͼ 60 human patients with a mean age of 43.6 and body mass index (BMI) of 27. Methods . We examined the ASC yield per unit volume of lipoaspirate tissue, the surface antigen profi le based on fl ow cytometry, histochemical differentiation potential along the adipogenic and osteogenic pathways, and expression of adipogenic mRNA by transcriptomic microarray and reverse transcription (RT) -polymerase chain reaction (PCR). Results . The population ( n ϭ 64) of predominantly Caucasian (84.3%) female (90.6%) donors had a mean age of 43.6 Ϯ 11.1 years and a mean BMI of 27.0 Ϯ 3.8. A yield of 375 Ϯ 142 ϫ 10 3 ASC was obtained per milliliter of lipoaspirate within a 4.1 Ϯ 0.7-day culture period ( n ϭ 62). The ASC population was uniformly CD29 ϩ CD34 ϩ CD44 lo CD45 lo CD73 ϩ CD90 ϩ CD105 ϩ and capable of undergoing both adipogenesis and osteogenesis in vitro based on Oil Red O and Alizarin Red staining, respectively. Adipogenic differentiation was associated with a signifi cant induction of multiple mRNA associated with lipid storage and synthesis based on microarray analysis of n ϭ 3 donors. During an adipogenic differentiation timecourse, representative mRNA (adiponectin, C/EBP α , leptin and LPL) displayed increases of several orders of magnitude. Conclusions . These fi ndings demonstrate the reproducibility of subcutaneous lipoaspirates as a consistent and abundant source of functional ASC from donors across a spectrum of ages and BMI. These results have relevance for regenerative medical applications exploiting autologous and allogeneic ASC for soft and hard tissue engineering.
2009
Adipose tissue is a dynamic endocrine organ with a central role in metabolism regulation. Functional differences in adipose tissue seem associated with the regional distribution of fat depots, in particular in subcutaneous and visceral omental pads. Here, we report for the first time the isolation of human adipose-derived adult stem cells from visceral omental and subcutaneous fat (V-ASCs and S-ASCs, respectively) from the same subject. Immunophenotyping shows that plastic culturing selects homogeneous cell populations of V-ASCs and S-ASCs from the corresponding stromal vascular fractions (SVFs), sharing typical markers of mesenchymal stem cells. Electron microscopy and electrophysiological and real-time RT-PCR analyses confirm the mesenchymal stem nature of both V-ASCs and S-ASCs, while no significant differences in a limited pattern of cytokine/chemokine expression can be detected. Similar to S-ASCs, V-ASCs can differentiate in vitro toward adipogenic, osteogenic, chondrogenic, muscular, and neuronal lineages, as demonstrated by histochemical, immunofluorescence, real-time RT-PCR, and electrophysiological analyses, suggesting the multipotency of such adult stem cells. Our data demonstrate that both visceral and subcutaneous adipose tissues are a source of pluripotent stem cells with multigermline potential. However, the visceral rather than the subcutaneous ASC could represent a more appropriate in vitro cell model for investigating the molecular mechanisms implicated in the pathophysiology of metabolic disorders such as obesity.
Cytotechnology, 2013
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.
Adipose tissue-derived stem cells show considerable promise for regenerative medicine applications
Cellular & Molecular Biology Letters, 2013
The stromal-vascular cell fraction (SVF) of adipose tissue can be an abundant source of both multipotent and pluripotent stem cells, known as adipose-derived stem cells or adipose tissue-derived stromal cells (ADSCs). The SVF also contains vascular cells, targeted progenitor cells, and preadipocytes. Stromal cells isolated from adipose tissue express common surface antigens, show the ability to adhere to plastic, and produce forms that resemble fibroblasts. They are characterized by a high proliferation potential and the ability to differentiate into cells of meso-, ecto- and endodermal origin. Although stem cells obtained from an adult organism have smaller capabilities for differentiation in comparison to embryonic and induced pluripotent stem cells (iPSs), the cost of obtaining them is significantly lower. The 40 years of research that mainly focused on the potential of bone marrow stem cells (BMSCs) revealed a number of negative factors: the painful sampling procedure, frequent ...
Journal of vascular surgery, 2016
One of the rate-limiting barriers within the field of vascular tissue engineering is the lengthy fabrication time associated with expanding appropriate cell types in culture. One particularly attractive cell type for this purpose is the adipose-derived mesenchymal stem cell (AD-MSC), which is abundant and easily harvested from liposuction procedures. Even this cell type has its drawbacks, however, including the required culture period for expansion, which could pose risks of cellular transformation or contamination. Eliminating culture entirely would be ideal to avoid these concerns. In this study, we used the raw population of cells obtained after digestion of human liposuction aspirates, known as the stromal vascular fraction (SVF), as an abundant, culture-free cell source for tissue-engineered vascular grafts (TEVGs). SVF cells and donor-paired cultured AD-MSCs were first assessed for their abilities to differentiate into vascular smooth muscle cells (SMCs) after angiotensin II s...
Use of Freshly Isolated Human Adipose Stromal Cells for Clinical Applications
Aesthetic Surgery Journal, 2017
The clinical use of adipose-derived cells is being explored very actively around the world for various human diseases. Adipose tissue is an abundant tissue source that can be easily harvested using liposuction. Human lipoaspirates contain a significant amount of mesenchymal stromal cells, as well as other progenitors and terminally differentiated cell types. This review covers the isolation of adipose stromal vascular fraction (SVF), the quality control and safety analysis of freshly isolated cell suspensions. The comparison between freshly isolated stromal cells and culture expanded cells from adipose tissue samples is also highlighted. This article provides a brief but comprehensive review about SVF isolation in the clinical setting, cell characterization, and biological potency of freshly obtained adipose stromal cells.