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Differentiation of Bone Marrow Mesenchymal Stem Cells into Hair Like Cells by Co-culture Method

Author: YangWenFei
Tutor: HuangJianMin
School: Fujian Medical
Course: Department of Otolaryngology - Head and Neck Surgery
Keywords: Bone marrow mesenchymal stem cells Hair cells Co-culture Differentiation
CLC: R764.43
Type: Master's thesis
Year: 2011
Downloads: 26
Quote: 0
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Abstract


Objective: To establish in vitro co-culture statutory differentiation to the hair cell -like cells to the induction of bone marrow mesenchymal stem cells (BMSCs) . Methods : whole bone marrow adherence method isolation and culture of mesenchymal stem cells in mouse bone marrow . And passaged and purified . The third alternative fluorescent immunohistochemical method to detect cell surface marker protein of CD44 , CD34 , CD106, Vimentin in expression . Mining newborn mouse brain glial cells (ASTs) in vitro cultivation to the third generation , the expression of the marker protein GFAP expression in glial cells using immunofluorescence assay . Mining newborn mouse ear basement membrane (BAMCs) cells in vitro amplification spare nurturing to the third generation . The third generation BMSCs co-cultured with ASTs , and combined with RA (DMEM/F12/RA 10? S) induced by 7 days of culture , the BMSCs differentiation into neural precursor cells . Induced cells with a fluorescent immunohistochemical method to detect neural precursor signs protein expression (Nestin) . The resulting cells after induction with BAMCs co-cultured in DMEM/F12 10 ? 14 days in S induce their differentiation to the hair cell -like cells . And use the fluorescent immunohistochemical method to detect markers of hair cells (Math1 MyosinVIIa) , protein . Math1 and MyosinVIIa mRNA expression by RT-PCR assay . Results: training and joint RA induced BMSCs expressing neural precursor cell-specific protein of Nestin ASTs total . The after such neural precursor cells co-cultured with BAMCs , some cells express the hair cell - specific marker protein the Math1, MyosinVIIa mRNA and protein . Conclusion : BMSCs were by co-culture with ASTs and joint RA induction resulting cells co-cultured with BAMCs part of the expression of Math1 and MyosinVIIa mRNA and the protein in the hair cell-like cells can be induced to differentiate . Co-culture method can induce BMSCs differentiation to the hair cell -like cells .

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CLC: > Medicine, health > Otorhinolaryngology > Otology,ear disease > Ear nervous system diseases > Deaf
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