Abstract
Objective: Placenta, umbilical cord, and blood are alternative sources of stem cells. We designed an experiment to isolate and culture stem cells from human amniotic membrane. And we made a plan to induce auditory hair cells and neurons from the stem cells of amniotic membrane.
Method: Human amniotic membrane was obtained.) We dissociated amniotic membrane with trypsin and transferred it to culture medium which contained 10ï¼… fetal bovine serum and 1ï¼… penicillin-streptomycin. We sub cultured it until the third generation was used for further experimental procedures. For differentiation into neural progenitor cells, we cultured amniotic membrane stem cells in EGF (epidermal growth factor) and β-FGF (fibrous growth factor) and for differentiation into hair cells and neurons, we cultured in neurobasal medium.
Results: We can isolate and culture stem cells from human amniotic membrane. By flow cytometric analysis of stem cells from human amniotic membrane, they showed positive responses at mesenchymal stem cell marker (CD 73, CD90, CD105) and negative responses at hematopoietic stem cell marker (CD34, CD14, CD19, HLA). The results of culture at different culture medium showed a differentiation to neural progenitor cells, hair cells, and neuron, and we confirmed by immunofluorescence staining and RT-PCR.
Conclusion: We demonstrated that amniotic membrane–derived stem cells could be differentiated into neuronal progenitor cells, and this could lead to the differentiation into hair cells and neurons.
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