Purpose:To study preliminarily in vitro induced differentiation of embryonic stem cells into neurons for further investigation of an alternative for the treatment of glaumatous neuropathy.Materials and methods:Supern...Purpose:To study preliminarily in vitro induced differentiation of embryonic stem cells into neurons for further investigation of an alternative for the treatment of glaumatous neuropathy.Materials and methods:Supernatant of cultured Buffalo rat liver cells(buffalorat liver cell-conditioned medium,BRL-CM)was used for culturing embryonic stem cells(ES-D3 cell line),Morphological features of undifferentiated EScells were studied by HE staining and electron microscopy.Based on the methods used by Bain et al,we modified the methods and used retinoic acid(RA)as an inducer to differentiate ES-D3 cells and cytosine arabinoside(Ara-C)as inhibitor of proliferative cells.The growth of the cells was observed under phase contrast microscope.Results:ES-D3cells cultured by BRL-CMgrew in aggregates and remained undifferentiated.Electromicroscopy showed large nucleus and a large amount of mitochondria in undifferentiated ES cells and many processes on the surfaces.In the first day after the adding of retinoic acid,some neuron-like cells with one,two or more processes were present.In the second day after adding RAand the first day after the plus of 10μm Ara-C,a large amount of neuron-like cells appeared,with the formation of neuron-like networks.Con clusions:Combined use of RA and Ara-Ccan induce ES cells to differentiate into neuron-like cells.Our present preliminary study might provide insights into an alternative for the treatment of glaucomatous neuropathy by the transplantation of embryonic stem cells.Eye Science2000;16:1-6.展开更多
基金This paper is granted by 973 Branched Project(G1999054301)National Natural Sciences Foundation of China (No.39770789)+1 种基金Natural Sciences Foundation of Guangdong Province (No.990092) 211 Project Foundation (No.98010)
文摘Purpose:To study preliminarily in vitro induced differentiation of embryonic stem cells into neurons for further investigation of an alternative for the treatment of glaumatous neuropathy.Materials and methods:Supernatant of cultured Buffalo rat liver cells(buffalorat liver cell-conditioned medium,BRL-CM)was used for culturing embryonic stem cells(ES-D3 cell line),Morphological features of undifferentiated EScells were studied by HE staining and electron microscopy.Based on the methods used by Bain et al,we modified the methods and used retinoic acid(RA)as an inducer to differentiate ES-D3 cells and cytosine arabinoside(Ara-C)as inhibitor of proliferative cells.The growth of the cells was observed under phase contrast microscope.Results:ES-D3cells cultured by BRL-CMgrew in aggregates and remained undifferentiated.Electromicroscopy showed large nucleus and a large amount of mitochondria in undifferentiated ES cells and many processes on the surfaces.In the first day after the adding of retinoic acid,some neuron-like cells with one,two or more processes were present.In the second day after adding RAand the first day after the plus of 10μm Ara-C,a large amount of neuron-like cells appeared,with the formation of neuron-like networks.Con clusions:Combined use of RA and Ara-Ccan induce ES cells to differentiate into neuron-like cells.Our present preliminary study might provide insights into an alternative for the treatment of glaucomatous neuropathy by the transplantation of embryonic stem cells.Eye Science2000;16:1-6.
