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Isolation characterization and culture of Thy1-positive cells from fetal rat livers

机译:胎鼠肝脏Thy1阳性细胞的分离鉴定和培养

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摘要

AIM: To investigate whether Thy1 recognizes oval cells in the fetal liver and to characterize the cultured Thy1- selected cells from E14 rat livers.METHODS: Thy1 populations were analyzed by fluorescence activated cell sorter analysis. Thy1 positive cells were isolated using magnetic beads. Hepatic markers were detected by Western blotting, immunocytochemistry and RT-PCR.RESULTS: The percentage of Thy1-positive cells decreased during early development of fetal rat liver (E13-E16). E14 fetal livers contained 7.8% Thy1 positive cells, of which 61% were positive for α-fetoprotein (AFP) and 25% expressed albumin. The Thy1+ population expressed oval cell markers c-Kit and CXCR4, liver enriched-transcription factors HNF1α and HNF6, hepatocytic markers albumin, AFP and cytokeratin 18, and biliary marker cytokeratin 19. Thy1- selected cells formed only mesenchymal colonies when plated on collagen and in serum-containing media. Thy1 selected cells were able to form hepatic colonies positive for HNF1α, HNF6, albumin, AFP, cytokeratin 18, cytokeratin 19 and glycogen, when grown on STO feeder layers in serum free-media.CONCLUSION: Oval cells positive for Thy1 are present in early liver embryonic stages.
机译:目的:研究Thy1是否能识别胎儿肝脏中的卵圆形细胞,并鉴定来自E14大鼠肝脏的培养的Thy1选择细胞。方法:通过荧光激活细胞分选仪分析Thy1群体。使用磁珠分离Thy1阳性细胞。结果:胎鼠肝脏早期发育(E13-E16)过程中,Thy1阳性细胞百分率下降,Western blotting,免疫细胞化学和RT-PCR检测肝标志物。 E14胎儿肝脏含有7.8%的Thy1阳性细胞,其中61%的α-甲胎蛋白(AFP)阳性和25%的白蛋白表达。 Thy1 +人群表达卵形细胞标志物c-Kit和CXCR4,肝富集转录因子HNF1α和HNF6,肝细胞标志物白蛋白,AFP和细胞角蛋白18以及胆汁标志物细胞角蛋白19。在含血清的培养基中。当在无血清培养基中的STO饲养层上生长时,选择的Thy1细胞能够形成HNF1α,HNF6,白蛋白,AFP,细胞角蛋白18,细胞角蛋白19和糖原阳性的肝菌落。肝胚胎期。

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