Transfection of adult canine Schwann cells and olfactory ensheathing cells at early and late passage with human TERT differentially affects growth factor responsiveness and in vitro growth
Transfection of adult canine Schwann cells and olfactory ensheathing cells at early and late passage with human TERT differentially affects growth factor responsiveness and in vitro growth
复制标题
DOI：
10.1016/j.jneumeth.2008.08.030
复制
发表时间：
2009-01-30
影响因子：
3
通讯作者：
Baumgaertner, Wolfgang
中科院分区：
文献类型：
--
作者：
Techangamsuwan, Somporn;Kreutzer, Robert;Baumgaertner, Wolfgang
成年犬雪旺细胞和嗅鞘细胞（OEC）是密切相关的细胞类型，被认为是神经修复转化研究的有吸引力的候选者。为了通过比较用于移植目的的永生化雪旺细胞和 OEC 的体外特性来建立可靠的细胞来源，我们用人端粒酶逆转录酶 (hTERT) 转染这两种细胞类型。异位 hTERT 表达已被证明可以诱导各种细胞类型的永生化，而不会显着改变其表型。从成年犬中分离雪旺细胞和 OEC，在早期 (P4) 和晚期 (P26) 传代时用 hTERT 转染，在存在和不存在成纤维细胞生长因子-2 (FGF-2) 的情况下表征体外增殖、抗原表达和衰老相关基因。用 FGF-2 处理但未用 FGF-2 处理的晚期神经胶质细胞中的异位 hTERT 表达可阻止在未转染细胞中观察到的增殖下降。永生化不改变p75(NTR)和GFAP，但改变04和A2B5表达。与此相反，早期传代 hTERT 转染显着降低了两种细胞类型中不依赖于 FGF-2 的增殖，并降低了 04 和 GFAP 的表达。转染不会改变衰老相关基因（例如 p53 和 p16）的 mRNA 表达。雪旺细胞和 OEC 之间没有发现实质性差异，强调了两种细胞类型的密切关系。总之，我们建立了成年犬雪旺细胞和 OEC 的稳定来源，并证明 hTERT 表达对体外生长和生长因子反应性的影响取决于复制年龄。 (C) 2008 Elsevier B.V. 保留所有权利。
Adult canine Schwann cells and olfactory ensheathing cells (OECs) are closely related cell types that are considered attractive candidates for translational studies of neural repair. To establish a reliable cell source by comparing the in vitro properties of immortalized Schwann cells and OECs for transplantation purposes, we transfected both cell types with human telomerase reverse transcriptase (hTERT). Ectopic hTERT expression has been shown to induce immortalization of various cell types without substantial alterations of their phenotypes. Schwann cells and OECs were isolated from adult dogs, transfected with hTERT at early (P4) and late passage (P26), characterized regarding in vitro proliferation, antigenic expression and senescence-associated genes in the presence and absence of fibroblast growth factor-2 (FGF-2). Ectopic hTERT expression in late passage glia treated with but not without FGF-2 prevented the decline in proliferation observed in non-transfected cells. Immortalization did not alter p75(NTR) and GFAP but 04 and A2B5 expression. Contrary to this, early passage hTERT transfection significantly reduced proliferation independent of FGF-2 and lowered expression of 04 and GFAP in both cell types. Transfection did not alter mRNA expression of senescence-associated genes such as p53 and p16. No substantial differences were found between Schwann cells and OECs underscoring the close relationship of both cell types. Taken together, we established a stable source of adult canine Schwann cells and OECs and demonstrated that the effects of hTERT expression on in vitro growth and growth factor responsiveness depend on the replicative age. (C) 2008 Elsevier B.V. All rights reserved.