Isabel Zvibel, Miri Bronstein, Einav Hubel, Ella Bar-Lev, Zamir Halpern, Ran Oren
Liver Unit, Gastroenterology Institute, Tel Aviv Sourasky Medical Center, Israel.
World J Gastroenterol. 2006 Jun 28;12(24):3841-7. doi: 10.3748/wjg.v12.i24.3841.
To investigate whether Thy1 recognizes oval cells in the fetal liver and to characterize the cultured Thy1- selected cells from E14 rat livers.
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.
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 alpha-fetoprotein (AFP) and 25% expressed albumin. The Thy1+ population expressed oval cell markers c-Kit and CXCR4, liver enriched-transcription factors HNF1alpha 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 HNF1alpha, HNF6, albumin, AFP, cytokeratin 18, cytokeratin 19 and glycogen, when grown on STO feeder layers in serum free-media.
Oval cells positive for Thy1 are present in early liver embryonic stages.
研究Thy1是否识别胎肝中的卵圆细胞,并对从E14大鼠肝脏中经Thy1筛选培养的细胞进行特性分析。
通过荧光激活细胞分选分析对Thy1群体进行分析。使用磁珠分离Thy1阳性细胞。通过蛋白质免疫印迹法、免疫细胞化学和逆转录聚合酶链反应检测肝脏标志物。
在胎鼠肝脏早期发育阶段(E13 - E16),Thy1阳性细胞的百分比下降。E14胎肝中含有7.8%的Thy1阳性细胞,其中61%的细胞甲胎蛋白(AFP)呈阳性,25%的细胞表达白蛋白。Thy1⁺群体表达卵圆细胞标志物c-Kit和CXCR4、肝脏富集转录因子HNF1α和HNF6、肝细胞标志物白蛋白、AFP和细胞角蛋白18,以及胆管标志物细胞角蛋白19。当接种在胶原蛋白上并置于含血清培养基中时,经Thy1筛选的细胞仅形成间充质集落。当在无血清培养基中的STO饲养层上生长时,经Thy1筛选的细胞能够形成对HNF1α、HNF6、白蛋白、AFP、细胞角蛋白18、细胞角蛋白19和糖原呈阳性的肝集落。
Thy1阳性的卵圆细胞存在于肝脏胚胎早期阶段。