论文部分内容阅读
目的:利用形态学研究方法,探讨神经干细胞特异性标志蛋白的表达和增殖分化特性,鉴定人类羊膜组织中神经干/前体细胞的存在。方法:利用免疫组织化学法,检测人类羊膜组织和培养人类羊膜细胞中nestin、musashi-1、vimentin和PSA-NCAM等神经干细胞特异性标志蛋白的表达,利用BrdU掺入实验鉴定羊膜组织来源神经干/前体细胞的增殖分化能力。结果:免疫组织化学检测证实人羊膜组织表达nestin、musashi-1、CD133和vimentin等神经干细胞特异性标志蛋白,主要分布于羊膜组织的间质层。培养羊膜细胞的免疫荧光化学染色显示nestin、musashi-1、vimentin和PSA-NCAM阳性细胞的存在。BrdU掺入的组织形态学实验显示培养羊膜细胞中存在BrdU标记阳性细胞,其中具有BrdU与nestin、musashi-1、vimentin双标阳性细胞,显示羊膜细胞中存在具有增殖活性神经干细胞标记蛋白表达阳性细胞;而BrdU与β-tubullinIII、TH和GFAP双标阳性细胞的存在,表明神经元和胶质细胞是由培养羊膜细胞增殖分化的。结论:羊膜组织中存在神经干细胞特异性标记蛋白Nestin、Musashi-1、CD133、PSA-NCAM和Vimentin表达阳性细胞,BrdU掺入的组织形态学结果不仅证实羊膜细胞中Nestin、Musashi-1、Vimentin阳性细胞具有增殖活性,而且其中存在已分化的神经元和胶质细胞,提示羊膜组织细胞内存在神经干/前体细胞。
OBJECTIVE: To investigate the expression, proliferation and differentiation of neural stem cell-specific marker proteins and to identify the existence of neural stem / progenitor cells in human amniotic membrane by morphological methods. Methods: The expressions of nestin, musashi-1, vimentin and PSA-NCAM were detected by immunohistochemistry in human amnion and cultured human amniotic cells. BrdU incorporation assay was used to identify neural stem cells derived from amniotic tissue Proliferation and differentiation of precursor cells. Results: Immunohistochemistry confirmed that neural stem cell-specific marker proteins such as nestin, musashi-1, CD133 and vimentin were expressed in human amniotic membrane, which mainly distributed in the interstitial layer of amniotic tissue. Immunofluorescence staining of cultured amniotic cells revealed the presence of nestin, musashi-1, vimentin, and PSA-NCAM positive cells. BrdU incorporation of the histomorphological experiments showed that there are BrdU positive cells in cultured amniotic cells, with BrdU and nestin, musashi-1, vimentin double-labeled positive cells, there is a presence of proliferating neural stem cell marker protein expression in amniotic cells positive cells ; While the presence of BrdU and β-tubullin III, TH and GFAP double positive cells, indicating that neurons and glial cells by cultured amniotic cells proliferation and differentiation. CONCLUSIONS: Neural stem cell-specific marker proteins Nestin, Musashi-1, CD133, PSA-NCAM and Vimentin are present in amniotic tissues. The histological results of BrdU incorporation not only confirm the positive expression of Nestin, Musashi-1 and Vimentin in amniotic cells Cells have proliferative activity, and there are differentiated neurons and glial cells, suggesting the presence of neural stem / precursor cells in amniotic tissue cells.