血小板衍生生长因子-C对人毛乳头细胞生物学活性的影响

Effect of PDGF-C on biological characters of human dermal papilla cells in vitro

目的观察血小板衍生生长因子-C（PDGF-C）对体外培养的人毛乳头细胞生物学作用的影响。方法取整形外科除皱患者所弃头皮分离培养毛乳头，免疫荧光细胞染色法鉴定碱性磷酸酶在人毛乳头细胞的表达，CCK-8法检测浓度分别为0、10、20、30、40ng／ml的PDGF—C，在0、1、2、3、4、5、6d7个时间点对体外培养毛乳头细胞增殖的影响，绘制细胞生长曲线。流式细胞仪分析最适合浓度下人毛乳头细胞的细胞周期变化。Transwell细胞迁移实验及细胞划痕实验测定PDGF—C对毛乳头细胞迁移、趋化能力。结果碱性磷酸酶在人毛乳头细胞中呈阳性表达。PDGF—C在0～40ng/ml范围内，能明显促进人毛乳头细胞的增殖，并成剂量依赖关系，对人毛乳头细胞的促增殖作用在30ng／ml达高峰，差异有统计学意义（P〈0．05），在PDGF-C为30ng／ml条件下体外培养的人毛乳头细胞，处于S期细胞的比例较对照组显著增加，差异有统计学意义（P〈0．05）。PDGF-C组相对于对照组向划痕处迁移细胞数明显增多。PDGF-C组向Transwell小室下面迁移细胞数多于对照组，2组比较差异有统计学意义（P〈0．05）。PDGF-C组碱性磷酸酶活性能力增高，与对照组比较差异有统计学意义（P〈0．05）。结论PDGF—C可促进体外培养的人毛乳头细胞的增殖、迁移及诱导能力。

Objective To determine the effect of platelet-derived growth factor-C （PDGF-C） on biological characters of human hair dermal papilla cells cultured in vitro. Methods The human dermal papilla cells（HDPCs） were isolated from human hair skin obtained from rhytidectomy procedure and then cultured in vitro. Cell counting and CCK-8 assay were used to detect the effects of PDGF-C （0,10,20,30 and 40ng/ml） on the proliferation of HDPCs at 0,1 d, 2d, 3d,4d, 5d, 6d. Under the optimal concentration, flow cytometry was used to detect cell phases. Transwell assay and cell scratch test were performed to detect cell migration. Alkaline Phosphatase Activity Assay kit was used to detect the inductive activity of HDPCs. Results PDGF-C significantly induced the proliferation of HDPCs. PDGF-C of 30ng/mL promoted the HDPCs proliferation at a summit and increased the percentage of the cells arrested at S phase （P 〈 0.05）. PDGF-C also increased the migration populations of cultured HDPCs. The cell number in lower side of the transwell insert membrane of 30ng/ml PDGF-C treated group was （361.3 ±24.95） while the control was （246.8 ±7. 525） ,showing significant difference （ P 〈 0.05 ）. The alkaline phosphate activity of cultured HDPCs was increased comparing to the control group, the difference was significant （ P 〈 0.05 ）. Conclusions PDGF-C can promote the proliferation, migration and inductive activity of cultured human dermal papilla cells, which might be beneficial to promote the cultivation of human dermal papilla cells in vitro.