Small hepatocytes in culture develop polarized transporter expression and differentiation
Small hepatocytes in culture develop polarized transporter expression and differentiation
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培养中的小肝细胞产生极化转运蛋白表达和分化
DOI：
10.1242/jcs.01279
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发表时间：
2004
影响因子：
4
通讯作者：
B. Stieger
中科院分区：
文献类型：
--
作者：
Marguerite;M. Höchli;D. Inderbitzin;P. Meier;B. Stieger
大鼠小肝细胞已被证明在培养物中增殖，并在体外与分化的肝细胞形成类器官。为了评估大鼠小肝细胞在培养9周期间极化转运蛋白分化的程度，我们研究了肝细胞主要胆汁盐和有机阴离子转运系统[即基底外侧牛磺胆酸钠共转运蛋白（Ntcp），有机阴离子转运多肽1b 2（Oatp 1b 2），小管胆盐输出泵（Bsep）和多药耐药相关蛋白2（Mrp 2）]。通过形态学、甲胎蛋白、白蛋白和Mrp 1表达评估，小肝细胞在培养物中增殖和分化，并形成界限分明的集落。极化表面转运蛋白的表达是明显的Ntcp，Oatp 1b 2和Mrp 2培养5周后，7周后Bsep。培养9周后，绝大多数成熟肝细胞在基底外侧表达Ntcp/Oatp 1b 2，在小管质膜区域表达Bsep/Mrp 2。这种极化转运蛋白的表达伴随着荧光素二乙酸酯和胆甘氨酰荧光素的小管分泌。此外，堆积的菌落内形成了一个立体化的胆小管网络。这些数据表明，培养的大鼠小肝细胞获得完全分化的转运蛋白表达表型在其发展成肝的成熟肝细胞的类器官样簇。因此，转运蛋白表达的时间依赖性序列反映了发育中大鼠肝脏转运蛋白表达的个体发生，支持小肝细胞对应于来自胚胎成肝细胞和/或来自不同库的“定向肝细胞祖细胞”的肝细胞谱系的概念。
Rat small hepatocytes have been shown to proliferate in culture and to form organoids with differentiated hepatocytes in vitro. To evaluate the degree of polarized transporter differentiation of rat small hepatocytes during 9 weeks of culturing, we studied the time-dependent expression and subcellular localization of the major bile salt and organic anion transport systems of hepatocytes [i.e. the basolateral sodium-taurocholate co-transporting protein (Ntcp), organic-anion-transporting polypeptide 1b2 (Oatp1b2), the canalicular bile-salt export pump (Bsep) and multidrug-resistance-associated protein 2 (Mrp2)]. Small hepatocytes proliferated and differentiated in culture and formed sharply demarcated colonies as assessed by morphology, α-fetoprotein, albumin and Mrp1 expression. Polarized surface transporter expression was evident after 5 weeks of culturing for Ntcp, Oatp1b2 and Mrp2, and after 7 weeks for Bsep. After 9 weeks in culture, the vast majority of matured hepatocytes expressed Ntcp/Oatp1b2 at the basolateral and Bsep/Mrp2 at the canalicular plasma-membrane domains. This polarized transporter expression was accompanied by canalicular secretion of fluorescein-diacetate and cholylglycyl-fluorescein. Furthermore, an anastomizing three-dimensional network of bile canaliculi developed within piling-up colonies. These data demonstrate that cultured rat small hepatocytes acquire a fully differentiated transporter expression phenotype during their development into hepatic `organoid-like' clusters of mature hepatocytes. Thereby, the time-dependent sequence of transporter expression mirrored the ontogenesis of transporter expression in developing rat liver, supporting the concept that small hepatocytes correspond to the hepatocyte lineage derived from embryonic hepatoblasts and/or from a different pool of `committed hepatocyte progenitor cells'.
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小鼠有机阴离子转运多肽-5（一种肾脏特异性有机阴离子转运蛋白）的克隆、表达和个体发育。
DOI：
10.1006/bbrc.2000.4072
发表时间：
2001
期刊：
Biochemical and biophysical research communications.
影响因子：
--
作者：
Choudhuri,S;Ogura,K;Klaassen,CD
通讯作者：
Klaassen,CD
DOI：
10.1073/pnas.90.2.435
发表时间：
1993-01-15
影响因子：
11.1
作者：
BOYER, JL;HAGENBUCH, B;MEIER, PJ
通讯作者：
MEIER, PJ
小鼠胚胎成肝细胞的体外选择性双潜能分化。
DOI：
--
发表时间：
1997
期刊：
The American journal of pathology.
影响因子：
--
作者：
Rogler,LE
通讯作者：
Rogler,LE
个体发育过程中肝细胞质膜糖蛋白调节的改变。
影响因子：
3.7
作者：
Petell,JK;Quaroni,A;Hong,WJ;Hixson,DC;Amarri,S;Reif,S;Bujanover,Y
通讯作者：
Bujanover,Y