Two populations of Thy1-positive mesenchymal cells regulate in vitro maturation of hepatic progenitor cells
Two populations of Thy1-positive mesenchymal cells regulate in vitro maturation of hepatic progenitor cells
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DOI：
10.1152/ajpgi.00241.2006
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发表时间：
2007-02-01
影响因子：
4.5
通讯作者：
Ikai, Iwao
中科院分区：
文献类型：
--
作者：
Kamo, Naoko;Yasuchika, Kentaro;Ikai, Iwao
我们先前报道了CD 49 f（+）Thy 1（-）CD 45（-）（CD 49 f阳性）胎肝祖细胞（HPC）的体外成熟由来自胎肝的Thy 1阳性间充质细胞支持。这些间充质细胞制备物含有两个群体，一个为立方形，另一个为纺锤形。在这项研究中，我们确定粘蛋白型跨膜糖蛋白gp 38可以区分立方细胞梭形细胞的免疫细胞化学。RT-PCR分析显示分离的CD 49 f（+/-）Thy 1（+）gp 38（+）CD 45（-）（gp 38阳性）细胞和CD 49 ff（+/-）Thy 1（+）gp 38（-）CD 45（-）（gp 38阴性）细胞之间存在差异，而两种细胞均表达间充质细胞标志物。与gp 38阳性细胞的共培养促进了CD 49 f阳性HPC的成熟，这是通过高碘酸-希夫（PAS）染色阳性来估计的，而与gp 38阴性细胞的共培养维持了PAS染色阴性的CD 49 f阳性HPC。通过与gp 38阳性细胞共培养，HPC上成熟肝细胞标志物，如酪氨酸氨基转移酶、色氨酸-2，3-双加氧酶和葡萄糖-6-磷酸酶的表达上调。此外，透射电子显微镜显示收购的成熟肝细胞的功能与gp 38阳性细胞共培养的HPC。这种对HPC成熟的影响被来自gp 38阴性细胞的条件培养基抑制。相比之下，通过HPC的溴脱氧尿苷掺入的上调证明了与gp 38阴性细胞共培养的增殖效应。总之，这些结果表明，由gp 38阳性细胞促进的HPC体外成熟可能会受到gp 38阴性细胞的抑制作用的反对，这可能会维持HPC的不成熟，增殖状态。
We previously reported that the in vitro maturation of CD49f(+)Thy1(-)CD45(-)(CD49f positive) fetal hepatic progenitor cells (HPCs) is supported by Thy1-positive mesenchymal cells derived from the fetal liver. These mesenchymal cell preparations contain two populations, one of a cuboidal shape and the other spindle shaped in morphology. In this study, we determined that the mucin-type transmembrane glycoprotein gp38 could distinguish cuboidal cells from spindle cells by immunocytochemistry. RT-PCR analysis revealed differences between isolated CD49f (+/-)Thy1(+)gp38(+)CD45(-)(gp38 positive) cells and CD49ff(+/-)Thy1(+)gp38(-)CD45(-)(gp38 negative) cells, whereas both cells expressed mesenchymal cell markers. The coculture with gp38-positive cells promoted the maturation of CD49f-positive HPCs, which was estimated by positivity for periodic acid-Schiff (PAS) staining, whereas the coculture with gp38-negative cells maintained CD49f-positive HPCs negative for PAS staining. The expression of mature hepatocyte markers, such as tyrosine aminotransferase, tryptophan-2,3-dioxygenase, and glucose-6-phosphatase, were upregulated on HPCs by coculture with gp38-positive cells. Furthermore, transmission electron microscopy revealed the acquisition of mature hepatocyte features by HPCs cocultured with gp38-positive cells. This effect on maturation of HPCs was inhibited by the addition of conditioned medium derived from gp38-negative cells. By contrast, the upregulation of bromodeoxyuridine incorporation by HPCs demonstrated the proliferative effect of coculture with gp38-negative cells. In conclusion, these results suggest that in vitro maturation of HPCs promoted by gp38-positive cells may be opposed by an inhibitory effect of gp38-negative cells, which likely maintain the immature, proliferative state of HPCs.