脂肪干细胞促进人表皮角质形成细胞创面模型愈合的研究

Promotive effect of adipose-derived stem cells on the wound model of human epidermal keratinocyte in vitro

目的探讨脂肪干细胞（ADSCs）促进人表皮角质形成细胞（HEKa）创面模型愈合的可能性。方法体外培养大鼠ADSCs（rADSCs）（n=10）。实验组为rADSCs与HEKa直接共培养组（n=10），对照组为在transwell培养板建立的rADSCs与HEKa间接共培养（间接组，n=8）和单纯HEKa培养组（单纯组，n=8）。刮擦生长融合成片的HEKa细胞制备创面。培养24、48、72h后，计数迁移到创面的HEKa细胞数量并计算创面愈合率，检测HEKa细胞吸收脱氧胸腺嘧啶核苷的放射活性以判定HEKa细胞的增殖活性。结果实验组、间接组、单纯组在伤后24h分别有（9．2±0．2）、（5．0±0．3）、（4．2±0．3）个细胞／高倍视野；48h分别有（58．5±0．4）、（26．5±0．3）、（20．7±0．5）个细胞／高倍视野；72h分别有（125．8±0．4）、（43．0±0．5）、（35．6±0．5）个细胞／高倍视野越过创缘进入创面，实验组均明显优于两对照组（P〈0．05）。三组创面愈合率在伤后72h分别为61．0％±3．0％、35．0％±2．5％、32．0％±2．1％，实验组均明显优于两对照组（P〈0．05）。脱氧胸腺嘧啶核苷掺入培养基法表明三组HEKa细胞的放射性强度为（1440±210）、（1050±280）、（1130±390）cpm／10^5细胞，实验组与两对照组之间差异有统计学意义（P〈0．05）。结论rADSCs通过直接接触促进HEKa细胞的分裂增殖和迁移。

Objective To investigate the migrating effect of adipose derived stem cells （ADSCs） on the wound model of human epidermal keratinocyte（HEKa）. Methods Rat ADSCs （rADSCs） were isolated and cultured （n = 10）, rADSCs were direct co-cultured with HEKa cells in experiment group （experimental group, n = 10）. In the control groups, rADSCs were indirect co-cuhured with HEKa cells in transwell chambe （ indirect group, n = 8） , or HEKa was cultured alone （ single group, n = 8 ）. Then confluent HEKa cells were scraped to establish a wound model under invert microscope. After scraped 24, 48, and 72 h, cell numbers of which migrated across the edge of the wound was measured, the rate of wound healing was calculated by using SigmaScan Pro 5 software, and the proliferating effect of rADSCs on HEKa were examined by incorporation of [3HI thymidine. Results The cells migrated across the edge of wound after 24 hours in experimental group, indirect group, and single group were （9. 2 ±0. 2）, （5.0 ±0. 3）, （4. 2 ± 0.3）, and were （58.5±0.4）, （26.5±0.3）, （20.7±0.5） 48 hours after, and were （125.8±0.4）, （43.0 ± 0. 5 ）, （35.6 ± 0. 5 ） cells/HP 72 hours after, respectively; the numbers were all significantly higher in experimental group than those in control groups （ P 〈 0. 05 ）. The rates of wound healing after scraped 72 hours were 61.0% ±3.0%, 35.0% ±2.5% and 32.0 ±2. 1% , the outcome in experimental group was significantly better than in the control groups （ P 〈 0. 05 ）. And the thymidine feeding displayed the proliferation of HEKa in the three groups were （ 1440 ± 210 ） , （ 1050 ± 280） and （ 1130 ± 390） epm/10^5 cell, and there was significant difference between the experimental and the control groups （ P 〈 0. 05）. Conclusions The rADSCs can promote the migration of HEKa by direct contact with it.