Culture of newborn monkey liver epithelial progenitor cells in chemical defined serum-free medium
Culture of newborn monkey liver epithelial progenitor cells in chemical defined serum-free medium
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化学成分确定的无血清培养基中培养新生猴肝上皮祖细胞
DOI：
10.1007/s11626-010-9325-z
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发表时间：
2010-06
期刊：
影响因子：
--
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中科院分区：
文献类型：
--
作者：
对来自非人类灵长类动物的肝脏祖细胞的研究将有助于更好地了解它们的人类同行。在这项研究中，恒河猴肝上皮祖细胞(MLEPC)是从一小片新生的肝脏在化学定义的无血清培养液中分离出来的。消化后的肝细胞在含钙的培养液中形成细胞聚集体。产生了两种类型的细胞聚集体：细长的梭形细胞和多边形上皮细胞。细长的梭形细胞表达为波形蛋白和短臂，在我们的培养中，它们在5天内消失。其余类型为表达细胞角蛋白7(CK7)、CK8、CK18、巢蛋白、CD49f和E-cad的小多边形上皮细胞，它们是肝干细胞的标志，但不表达α-胎儿蛋白、白蛋白和CK19。如果在层粘连蛋白或鼠尾胶原胶上，它们可以增殖并传代，以启动集落。地塞米松和抑瘤素M可诱导分化细胞表达成熟的肝细胞标志物，如α-1抗胰蛋白酶、胞浆内糖原储存、吲哚青绿摄取和脂滴生成。如果将其转移到小鼠胚胎成纤维细胞饲养器上，可获得CK19阳性的胆管细胞，并形成甜甜圈样结构。因此，具有双功能的mLEPC来自新生的猴肝脏，可以作为评估人类细胞治疗的临床前模型。
Studies with hepatic progenitor cells from non-human primates would allow better understanding of their human counterparts. In this study, rhesus monkey liver epithelial progenitor cells (mLEPCs) were derived from a small piece of newborn livers in chemical defined serum-free medium. Digested hepatic cells were treated in Ca2+-containing medium to form cell aggregates. Two types of cell aggregates were generated: elongated spindle cells and polygonal epithelial cells. Elongated spindle cells were expressed as vimentin and brachyury, and they were disappeared within 5 d in our cultures. The remaining type consisted of small polygonal epithelial cells that expressed cytokeratin 7 (CK7), CK8, CK18, nestin, CD49f, and E-cad, the markers of hepatic stem cells, but were negative for α-fetoprotein, albumin, and CK19. They can proliferate and be passaged, if on laminin or rat tail collagen gel, to initiate colonies. When cultured with dexamethasone and oncostatin M, the expression of mature hepatocyte markers, such as α-1-antitrypsin, intracytoplasmic glycogen storage, indocyanine green uptake, and lipid droplet generation, were induced in differentiated cells. If transferred onto mouse embryonic fibroblasts feeders, they gave rise to CK19-positive cholangiocytes with formation of doughnut-like structure. Thus, mLEPCs with bipotency were derived from newborn monkey liver and may serve as a preclinical model for assessment of cell therapy in humans.
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DOI：
10.13039/501100004587
发表时间：
1983-08
期刊：
--
影响因子：
--
作者：
Cristina Rodríguez;Tomás Sobrino;José Carlos Gómez Sánchez;Angeles Almeida
通讯作者：
Cristina Rodríguez;Tomás Sobrino;José Carlos Gómez Sánchez;Angeles Almeida
DOI：
10.1083/jcb.200108066
发表时间：
2002-01-07
期刊：
The Journal of cell biology
影响因子：
--
作者：
Suzuki A;Zheng YW;Kaneko S;Onodera M;Fukao K;Nakauchi H;Taniguchi H
通讯作者：
Taniguchi H
影响因子：
4
作者：
Arends, Brigitte;Spee, Bart;Rothuizen, Jan
通讯作者：
Rothuizen, Jan
DOI：
10.3109/00498259409043224
发表时间：
1994-02
期刊：
Xenobiotica; the fate of foreign compounds in biological systems
影响因子：
--
作者：
H. Koebe;S. Pahernik;P. Eyer;F. Schildberg
通讯作者：
H. Koebe;S. Pahernik;P. Eyer;F. Schildberg
影响因子：
2.9
作者：
DUNN, JCY;TOMPKINS, RG;YARMUSH, ML
通讯作者：
YARMUSH, ML