Improving the neuronal differentiation efficiency of umbilical cord blood-derived mesenchymal stem cells cultivated under appropriate conditions (original) (raw)
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Rafidain Journal of Science
Human umbilical cord blood derived mesenchymal stem cells (MSCs) are regarded as valuable source for cell transplantation and cell therapy. These cells in their undifferentiated state are fibroblast-like in morphology, and these cells when treated with retinoic acid, or epidermal growth factor, rapidly assumed the morphology of multipolar neurons. The resulting differentiated cells express nestin marker, a neurofilament protein that is one of the most specific markers of multipotent neural stem cells. In conclusion, the present findings support the hypothesis that cord blood contains MSCs that can be induced to differentiate into neuronal pathway and express neuronal marker.
Journal of Neuroscience Research, 2008
We investigated the neurogenic potential of full-term human umbilical cord blood (hUCB)-derived multipotent mesenchymal stem cells (MSCs) in response to neural induction media or coculture with rat neural cells. Phenotypic and functional changes were assessed by immunocytochemistry, RT-PCR, and whole-cell patchclamp recordings. Naive MSCs expressed both mesodermal and ectodermal markers prior to neural induction. Exposure to retinoic acid, basic fibroblast growth factor, or cyclic adenosine monophosphate (cAMP) did not stimulate neural morphology, whereas exposure to dibutyryl cAMP and 3-isobutyl-1-methylxanthine stimulated a neuron-like morphology but also appeared to be cytotoxic. All protocols stimulated increases in expression of the neural precursor marker nestin, but expression of mature neuronal or glial markers MAP2 and GFAP was not observed. Nestin expression increases were serum level dependent. Electrophysiological properties of MSCs were studied with whole-cell patchclamp recordings. The MSCs possessed no ionic currents typical of neurons before or after neural induction protocols. Coculture of hUCB-derived MSCs and rat neural cells induced some MSCs to adopt an astrocytelike morphology and express GFAP protein and mRNA. Our data suggest hUCB-derived MSCs do not transdifferentiate into mature functioning neurons in response to the above neurogenic protocols; however, coculture with rat neural cells led to a minority adopting an astrocytelike phenotype. V
Many neural disorders are characterized by the loss of one or several types of neural cells. Human umbilical cord-derived mesenchymal cells (hUCMs) are capable of differentiating into neuron, astroglia-like and oligodendrocyte cell types. However, a reliable means of inducing the selective differentiation of hUCMs into neural cells in vitro has not yet been established. For induction of neural differentiation, hUCMs were seeded onto sterile glass slides and six various cocktails using a base medium (DMEM/LG) supplemented with 10 % FBS, retinoic acid (RA), dimethyl sulfoxide (DMSO), epidermal growth factor (EGF) and fibroblast growth factor (FGF) were used to compare their effect on neuronal, astrocyte and oligodandrocyte differentiation. The
Characterisation and Neurogenic Potential of Stem Cells from the Human Umbilical Cord Matrix
2010
Neural stem cell grafts can potentially repair damage or degeneration of the human central nervous system (CNS). However, the only neural cells reported to provide any benefit to date have been immature neural precursors derived from aborted foetuses. Recent studies suggest that neural cells can be derived from non-neural and non-embryonic tissues such as bone marrow, peripheral and umbilical cord blood, and umbilical cord matrix (Wharton's jelly). These tissues may therefore represent a more accessible source of cells for therapeutic repair and regeneration of the brain and spinal cord. Furthermore, they could potentially be obtained and grafted autologously, thereby reducing the risk of tissue rejection. At present, little is known about the origin, frequency and phenotypic characteristics of stem cells from the umbilical cord matrix. This aim of this study was to characterise and analyse the neurogenic potential of a potentially novel source of mesenchymal stem cells (MSCs) d...
Neural differentiation of human umbilical cord mesenchymal stem cells by cerebrospinal fluid
Iranian journal of child neurology, 2015
Wharton's jelly (WJ) is the gelatinous connective tissue from the umbilical cord. It is composed of mesenchymal stem cells, collagen fibers, and proteoglycans. The stem cells in WJ have properties that are interesting for research. For example, they are simple to harvest by noninvasive methods, provide large numbers of cells without risk to the donor, the stem cell population may be expanded in vitro, cryogenically stored, thawed, genetically manipulated, and differentiated in vitro. In our study, we investigated the effect of human cerebrospinal fluid (CSF) on neural differentiation of human WJ stem cells. The cells in passage 2 were induced into neural differentiation with different concentrations of human cerebrospinal fluid. Differentiation along with neural lineage was documented by expression of three neural markers: Nestin, Microtubule-Associated Protein 2 (MAP2), and Glial Fibrillary Astrocytic Protein (GFAP) for 21 days. The expression of the identified genes was confir...
American Journal of Bioscience and Bioengineering, 2015
Background: Mesenchymal stem cells (MSCs) are multipotent adult stem cells present in all tissues. They are present in bone marrow, and can differentiate in vitro into neurons, glial cells and myofibroblasts. MSCs have been proposed as sources of stem cells for regeneration of the CNS. Thus, one of the goals of regenerative medicine is to regenerate damaged brain tissue and spinal cord by harnessing the power of stem cells to initiate neurogenesis in damaged areas of the brain. Objective: The aim of this work is to study in-vitro induced neurogenesis using MSCs as model of stem cells. Methodology : Bone marrow-MSCs were isolated, expanded and passaged. MSCs were identified using morphology and flowcytometric analysis. Co-expression of Oct ¾ was done. MSCs were induced to neural lineage using Neural Induction Media (NIM) : a cocktail of retinoic acid dissolved in DEMSO, recombinant human Fibroblast Growth Factor (FGF) basic, recombinant human Epidermal Growth Factor (EGF) and Insulin-like Growth Factor I (IGF-I). Neural induction was verified morphologically, and immunologically using GFAP positivity and nestin expression. Results: BM-MSCs express CD44 and OCT ¾ which decrease with age. MSCs induced with NIM show morphological changes consistent with neurogenesis, positive GFAP and nestin expression as compared to the uninduced cells. Conclusion: MSCs isolated from bone marrow aspirate and can be differentiated into GFAP positive neural cells.
Biochemical and Biophysical Research Communications, 2017
Plasticity and developmental capacity of stem cells have now been established as a promising tool to restore the degenerative disorders. The linearity differentiation of human mesenchymal stem cells (hMSCs) into adipogenic, chondrogenic, osteogenic and even in neuronal subtypes has been demonstrated. The number of xenobiotics such as dexamethasone, insulin, isobutyl 1-methyle xanthine and retinoic acid has been reported for the potential to differentiate hMSCs into neuronal subtypes. But, the applicability of indigenous neurotrophic factor-nerve growth factor (NGF) has not been explored for the purpose. Thus, the present investigations were carried out to study the NGF induced neuronal differentiation of hMSCs. Following the isolation, purification and characterization of hMSCs were allowed to differentiate into neuronal subtypes under the influence of NGF (50 ng/mL). At various concentrations of NGF, the neuronal makers were analysed at both mRNA and protein levels. Cells, exposed with NGF were showing the significant and gradual increase in the neuronal markers in differentiating cells. The magnitude of expression of markers was maximum at day 4 of differentiation. NGF at 50 ng/mL concentration was found to induce neuronal differentiation of hMSCs into neuronal subtypes.
Multiple Sclerosis (MS) is an autoimmune inflammatory demyelinating disease of the central nervous system. The aim of this study was to investigate the neuroprotective effects of transplanted human umbilical cord blood mesenchymal stromal cells (UCB-MSC) derived neural progenitor cell (MDNPC) in EAE, an experimental model of MS. To initiate neuronal differentiation of UCB-MSCs, the pre-induction medium was removed and replaced with induction media containing retinoic acid, b FGF, h EGF, NGF, IBMX and ascorbic acid for one week. The expression of neural genes was examined in comparison to control group by real-time PCR assay. Then, experimental autoimmune encephalitis (EAE) was induced using myelin oligodendrocyte glycoprotein (MOG, 35-55 peptides) in 24 C57BL/6 mice. After induction, the mice were divided in four groups (n=6) as follows: healthy, PBS, UCB-MSCs and MDNPC, respectively. At the end of the study, disease status in all the groups was analyzed using hematoxylin-eosin (H&E) staining of brain sections. We found that UCB-MSCs exhibit neuronal differentiation potential in vitro and transplanted MDNPC lowered clinical score and reduced CNS leukocyte infiltration compared to untreated mice. Our results showed that MDNPC from UCB may be a proper candidate for regenerative therapy in MS and other neurodegenerative diseases.
Bulletin of Experimental Biology and Medicine, 2009
Human bone marrow MSC cultured in neurogenic medium containing EGF and FGFb demonstrated alteration of the phenotype and expression of neuronal precursor/early neuron markers nestin and NSE. Signals of expression of neuronal and oligodendroglial markers MAP-2, dm-20, and MBP were detected after prolongation of incubation in neurogenic medium to 2 weeks. Cells with neuronal morphology were immunopositive to early neuronal marker β-III-tubulin. Replacement of neurogenic medium for α-MEM with 10% fetal calf serum induced reversion of the phenotype to that typical for human MSC. This indicates high plasticity of the phenotype and expression profile of neuronal markers in MSC cultured under neurogenic conditions or possibility of dedifferentiation of MSC reaching the stage of neuronal precursors/early neurons.
Biochemical and Biophysical Research Communications, 2009
We have previously demonstrated that lineage negative cells (Lin neg ) from umbilical cord blood (UCB) develop into multipotent cells capable of differentiation into bone, muscle, endothelial and neural cells. The objective of this study was to determine the optimal conditions required for Lin neg UCB cells to differentiate into neuronal cells and oligodendrocytes. We demonstrate that early neural stage markers (nestin, neurofilament, A2B5 and Sox2) are expressed in Lin neg cells cultured in FGF4, SCF, Flt3-ligand reprogramming culture media followed by the early macroglial cell marker O4. Early stage oligodendrocyte markers CNPase, GalC, Olig2 and the late-stage marker MOSP are observed, as is the Schwann cell marker PMP22. In summary, Lin neg UCB cells, when appropriately cultured, are able to exhibit characteristics of neuronal and macroglial cells that can specifically differentiate into oligodendrocytes and Schwann cells and express proteins associated with myelin production after in vitro differentiation.