Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan.
Gene Ther. 2012 Jan;19(1):86-93. doi: 10.1038/gt.2011.62. Epub 2011 May 19.
The present study investigates the efficacy of polyethylenimine (PEI)-DNA complex that expressed human telomerase reverse transcriptase (hTERT) to transfect hair follicle stem cells and produce sufficient hTERT to stimulate hair growth. Transfection with pLC-hTERT-DNA-PEI complex (D+P group) in vitro induced expression of proliferating cell nuclear antigen in 35.8% of the purified stem cell population, suggesting enhanced cell proliferation. In vivo transfection efficiency of rat dorsal skin was determined by staining for β-gal activity. Cells positive for β-gal were located in the bulge region and dermal sheath of hair follicles. The follicles in the hTERT-transfected region entered anagenon day 15 after transfection, whereas non-transfected (Neg) controls remained in telogen. The similar effect was observed in 50-day-old rat dorsal skin. D+P group displayed a specific expression of hTERT and sufficient to initiate a transition to the anagen phase and promote new hair synthesis 18 days after the transfection. hTERT promoted follicle neogenesis following wounding. In all, 60 days after wounding, tissues of the D+P group showed more newly regenerating hair follicles (83±52 regenerated follicles per rat) in contrast to control group tissues (15±15 regenerated follicles per rat). These studies provide a potential approach for gene therapy of skin disease.
本研究旨在探讨转染人端粒酶逆转录酶(hTERT)的聚乙稀亚胺(PEI)-DNA 复合物对毛囊干细胞的转染效果,以产生足够的 hTERT 来刺激毛发生长。体外转染 pLC-hTERT-DNA-PEI 复合物(D+P 组)后,纯化的干细胞群体中有 35.8%表达增殖细胞核抗原,提示细胞增殖增强。通过β-半乳糖苷酶活性染色检测大鼠背部皮肤的体内转染效率。β-半乳糖苷酶阳性细胞位于毛囊的隆突区和真皮鞘。转染后 15 天,hTERT 转染区域的毛囊进入生长期,而非转染(Neg)对照组仍处于休止期。在 50 日龄大鼠背部皮肤中也观察到类似的效果。D+P 组 hTERT 的特异性表达足以启动向生长期的转变,并在转染后 18 天促进新的毛发生成。hTERT 促进了创伤后毛囊的新生。在创伤后 60 天,D+P 组组织显示出更多新再生的毛囊(每只大鼠 83±52 个再生毛囊),而对照组组织仅为 15±15 个再生毛囊。这些研究为皮肤病的基因治疗提供了一种潜在的方法。