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In-vitro Induction and Differentiation of Retinal Stem Cells and Mesenchymal Stem Cells into Retinal Neuro-epithelial Cells

Author: ZhangHui
Tutor: DongFangTian
School: Beijing Union Medical College
Course: Clinical
Keywords: retinal stem cells(RSCs) adipose mesenchymal stem cells(AMSCs) in-vitro induction retinal ganglion cells(RGCs) photo receptor
CLC: R774.1
Type: PhD thesis
Year: 2009
Downloads: 10
Quote: 0
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Abstract


Purpose:To explore and compare the in-vitro induction for differentiation of retinal stem cells (RSCs) and human adipose mesenchymal stem cells (AMSCs) into retinal neuro-epithelial cells.Methods:PN5rat retinal cells were digested by trypsinase and cultured by using their adherent ability to the wall of culture flask in DMEM/F12medium supplemented with10%FBS. The cells were purified by the method of continual passage. The differentiated RGCs were used as cell environment to induce the differentiation of AMSCs with EGF+bFGF+B27in DMEM/F12medium. RSCs were cultured by suspension method. RSCs and AMSCs were induced by EGF and Taurine. Immunofluorescence and RT-PCR were used to identify the differentiation of the cells.Results:In PN5rat retinal cells, there were mature RGCs, photoreceptors and Muller glia. RSCs, which expressed Nestin and Masuhi, were proved to exist in the PN5rat retinal cells. After induction with10%FBS, the cells turned into the same morphology and expressed Thy-1, Tuj1and Nogo-R. So the protein and mRNA of the induced cells proved to be the phenotypes of RGCs. After induction of the isolated RGCs, AMSCs had some typical morphology changes of neurons and expressed Tuj1and Rhodopsin on protein level, and expressed Nestin on mRNA level. With the induction of EGF and taurine, RSCs expressed Thy-1, Tuj1and GS. AMSCs expressed the neuron precursor marker Tuj1and glial marker GS by the induction of EGF.Conclusion:The components of the isolated PN5rat retinal cells are consistent with the development of PN5retina. There remain RSCs in PN5rat retina cells. After the induction of10%FBS, the cells can get the phenotypes of RGCs. AMSCs can express neural precursor marker and photoreceptor marker by the induction of purified RGCs. EGF and Taurine have positive effects on the differentiation of RSCs into retinal neuro-epithelial cells. And the effect of EGF on the differentiation of AMSCs is more prominent. RSCs can be induced to differentiate into retinal neuro-epithelial cells more easily than AMSCs. This study provides in-vitro induction evidence for stem cells as retinal implant cell donors.

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CLC: > Medicine, health > Ophthalmology > Retina and optic nerve diseases > Retinal disease
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