Cell dynamics and differentiation of conditionally immortalized human intestinal epithelial cells.
Cell dynamics and differentiation of conditionally immortalized human intestinal epithelial cells.
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DOI：
10.1053/gast.1997.v113.pm9322515
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发表时间：
1997-10
期刊：
影响因子：
29.4
通讯作者：
Andrea Quaroni;Jean-François Beaulieu
中科院分区：
文献类型：
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作者：
Andrea Quaroni;Jean-François Beaulieu
背景与目的肠上皮细胞系在单层培养中很难获得，这促使了新方法的使用，包括用癌基因或肿瘤病毒永生化。本研究的目的是获得条件永生化的人肠上皮细胞（温度敏感的胎儿人肠[tsFHI]细胞），并研究其生长和分化能力。方法用温度敏感的猴病毒40大肿瘤抗原转化人胎儿肠细胞，克隆，并筛选肠标志物的表达。结果tsFHI细胞的建立主要依赖于基于OptiMEM的生长培养基。在32 ℃时，tsFHI细胞增殖并显示隐窝细胞标志物。仅观察到少数分散的分化细胞以及显示凋亡的形态学标志的细胞。温度升高到39 ℃会导致不可逆的生长停滞和肠细胞样表型的获得。细胞周期蛋白依赖性激酶抑制剂p21/WAF 1/Cip 1的表达在转移至39 ℃的数小时内诱导，在刷状缘酶相应增加之前1-2天，表明在诱导中而不是在维持分化表型中的作用。结论：tsFHI细胞概括了体外肠细胞动力学的关键方面，似乎特别适合于研究细胞分化的早期事件。
BACKGROUND & AIMS Intestinal epithelial cell lines capable of differentiating in monolayer culture have been difficult to obtain, prompting the use of novel approaches including immortalization with oncogenes or tumor viruses. The aim of this study was to obtain conditionally immortalized human intestinal epithelial cells (temperature-sensitive fetal human intestinal [tsFHI] cells) and study their growth and differentiation capabilities. METHODS Intestinal cells from human fetuses were transformed with a temperature-sensitive simian virus 40 large tumor antigen, cloned, and screened for expression of intestinal markers. RESULTS Establishment of tsFHI cells was crucially dependent on a growth medium based on OptiMEM. At 32 degrees C, the tsFHI cells proliferate and display crypt cell markers. Only a few scattered differentiated cells are observed together with cells displaying the morphological signs of apoptosis. A shift to 39 degrees C results in irreversible growth arrest and acquisition of an enterocyte-like phenotype. Expression of the cyclin-dependent kinase inhibitor p21/WAF1/Cip1 is induced within hours of a shift to 39 degrees C 1-2 days before corresponding increases in brush border enzymes, suggesting a role in the induction rather than in the maintenance of a differentiated phenotype. CONCLUSIONS The tsFHI cells recapitulate key aspects of intestinal cell dynamics in vitro and seem particularly suited for the study of early events in cell differentiation.