Radiation Induces Diffusible Feeder Cell Factor(s) That Cooperate with ROCK Inhibitor to Conditionally Reprogram and Immortalize Epithelial Cells
Radiation Induces Diffusible Feeder Cell Factor(s) That Cooperate with ROCK Inhibitor to Conditionally Reprogram and Immortalize Epithelial Cells
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DOI：
10.1016/j.ajpath.2013.08.009
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发表时间：
2013-12-01
影响因子：
6
通讯作者：
Liu, Xuefeng
中科院分区：
文献类型：
--
作者：
Palechor-Ceron, Nancy;Suprynowicz, Frank A.;Liu, Xuefeng
饲养层细胞和Rho激酶抑制都是人上皮细胞条件重编程和永生化所必需的。在本研究中，我们证明了Rho激酶抑制剂Y-27632可以显著抑制角质形成细胞的分化，延长角质形成细胞在含血清培养中的寿命，但在没有饲养层细胞的情况下不会导致永生化。使用Transwell培养板，我们进一步证明了饲养层细胞和角质形成细胞之间的物理接触不是诱导永生化所必需的，更重要的是，这种诱导需要饲养层细胞的照射。与这些实验一致的是，条件培养液被证明可以诱导和维持条件永生化细胞，这伴随着端粒酶表达的增加。条件培养液的活性与辐射诱导饲养层细胞的凋亡有直接关系。因此，条件性重编程细胞的诱导是由Y-27632和由凋亡的饲养细胞释放的扩散因子(或多个因子)的组合所介导的。
Both feeder cells and Rho kinase inhibition are required for the conditional reprogramming and immortalization of human epithelial cells. In the present study, we demonstrated that the Rho kinase inhibitor Y-27632, significantly suppresses keratinocyte differentiation and extends life span in serum-containing medium but does not Lead to immortalization in the absence of feeder cells. Using Transwell culture plates, we further demonstrated that physical contact between the feeder cells and keratinocytes is not required for inducing immortalization and, more importantly, that irradiation of the feeder cells is required for this induction. Consistent with these experiments, conditioned medium was shown to induce and maintain conditionally immortalized cells, which was accompanied by increased telomerase expression. The activity of conditioned medium directly correlated with radiation-induced apoptosis of the feeder cells. Thus, the induction of conditionally reprogrammed cells is mediated by a combination of Y-27632 and a diffusible factor (or factors) released by apoptotic feeder cells.