Differentiation Of Bone Marrow Mesenchymal Stem Cells Into Hair Like Cells By Co-culture Method

Objective: To investigate a method of efficient production of hair like cells from bone marrow mesenchymal stem cells(BMSCs)in vitro.Methods: Mice bone marrow mesenchymal stem cells were separated and expanded by adherenee culture. We pass and depurate this cells to third generation.The surface molecule expressions of cells were detected by immunofluorescence. include CD44,CD34,CD106and Vimentin proteinum. The astrocytes were cultured in the method of different adherent time. We pass and depurate this cells to third generation. And detected the surface molecule of GFAP proteinum. Basement membrane (containing the organ of Corti) were isolated from postnatal day 1 mouse. Basement membrane cells were collected by mechanical dissociation and digested by 0.25% trypsin from the basement membrane.We used collected BMSCs co-cultured with astrocytes in DMEM/10%FBS+RA for 7 days, and induced BMSCs to differentiate into neural stem cell like cells.Examination the surface molecule of Nestin protein by immunofluorescence, Afterward,Take this cells co-cultured with basement membrane cells (BAMCs) in DMEM/10%FBS for 14 days.Checked the expression of cochlear hair cell markers (MyosinVIIa and Math1 mRNA and proteinum)in these differentiated cells .Results:BMSCs which were co-cultured with astrocytes for 7 days can become neuron like cells.then co-cultured with BAMCs for 14 days mRNA and proteinum of hair cell markers (Math1 and MyosinVIIa) were expressed. By using this way BMSCs can be induced to hair like cells.Conclusions: BMSCs can be induced to differentiate into neural stem like cells and hair like cells. co-culture method can induce BMSCs to differentiate into hair like cells.