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首页> 外文期刊>Cytokine >Liver sinusoidal endothelial cells support the survival and undifferentiated growth of the CGR8 mouse embryonic stem cell line: possible role of leukemia inhibitory factor (LIF).
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Liver sinusoidal endothelial cells support the survival and undifferentiated growth of the CGR8 mouse embryonic stem cell line: possible role of leukemia inhibitory factor (LIF).

机译:肝窦样内皮细胞支持CGR8小鼠胚胎干细胞系的存活和未分化生长:白血病抑制因子(LIF)的可能作用。

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摘要

Murine embryonic stem cells (muESC) are maintained and expanded in vitro by culturing in the presence of leukemia inhibitory factor (LIF) or by coculturing on murine embryonic fibroblast (MEF). Previously we have shown that liver sinusoidal endothelial cells (LSEC) promote the survival, proliferation and differentiation of hematopoietic stem cells. In the present study we investigated whether LSEC might promote the survival and undifferentiated growth of muESC. For these purposes, muESC (CGR8 cell line) were cultured on LSEC monolayers (muESC/LSEC) or in the presence of conditioned medium from LSEC cultures (muESC/LSEC-CM), both in the absence of LIF. Microscopic observation showed the growth of undifferentiated ESC colonies in both muESC/LSEC or muESC/LSEC-CM cultures. A significant reduction in the growth of undifferentiated ESC colonies was observed when ESC were cultured in LSEC-CM previously incubated with anti-LIF. RT-PCR and Western blot analysis showed that LSEC constitutively express LIF at the mRNA and protein level. At different times of culture, muESC were harvested and analyzed for the expression of embryonic markers (SSEA-1 and Oct-4) and differentiation capacity. Flow cytometry analysis showed the presence of a higher percentage of muESC (>90%) expressing SSEA-1 in muESC/LSEC-CM, as compared with muESC/LSEC cocultures. muESC obtained from both types of cultures formed embryoid bodies in vitro, and form teratomas in testicles of mice. These results provide the first evidence that LSEC support the in vitro survival, self-renewal, undifferentiated growth and differentiation capacity of the muESC CGR8 cell line.
机译:通过在白血病抑制因子(LIF)存在下培养或在鼠胚成纤维细胞(MEF)上共培养,鼠胚干细胞(muESC)得以在体外维持和扩增。以前我们已经表明,肝窦内皮细胞(LSEC)促进造血干细胞的存活,增殖和分化。在本研究中,我们调查了LSEC是否可能促进muESC的存活和未分化生长。为了这些目的,在不存在LIF的情况下,在LSEC单层(muESC / LSEC)上或在来自LSEC培养物的条件培养基(muESC / LSEC-CM)存在下培养muESC(CGR8细胞系)。显微镜观察显示在muESC / LSEC或muESC / LSEC-CM培养物中未分化的ESC集落的生长。当在预先与抗LIF孵育的LSEC-CM中培养ESC时,观察到未分化的ESC菌落的生长显着减少。 RT-PCR和蛋白质印迹分析表明LSEC在mRNA和蛋白水平上组成型表达LIF。在不同的培养时间,收集muESC并分析其胚胎标记(SSEA-1和Oct-4)的表达和分化能力。流式细胞仪分析表明,与muESC / LSEC共培养相比,在muESC / LSEC-CM中存在更高百分比的muESC(> 90%)表达SSEA-1。从两种类型的培养物中获得的muESC在体外形成胚状体,并在小鼠睾丸中形成畸胎瘤。这些结果提供了第一个证据,表明LSEC支持muESC CGR8细胞系的体外存活,自我更新,未分化的生长和分化能力。

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