表皮细胞培养上清中ET-1和SCF含量的检测

发布时间：2018-10-15 11:53

【摘要】：目的观察表皮细胞在3种培养基中的生长情况,检测不同时间点培养基中内皮素-1(ET-1)和干细胞因子(SCF)的水平。方法采用3种培养基培养表皮细胞,分别在培养24h,48h,72h,96h时收集培养上清,采用酶联免疫吸附试验(ELISA)方法检测ET-1和SCF水平,同时比较不同培养基中细胞的生长状态。结果三组培养上清中,培养基2(M2)中上清ET-1的含量最高;随着时间延长ET-1含量均增加,三组间比较差异有统计学意义(P均0.01);三组培养上清中SCF含量在第24~48h显著下降,三者之间的差异有统计学意义(P均0.01)。换液后ET-1和SCF变化趋势基本同换液前(P均0.01)。M2中细胞增殖最快,角质形成细胞呈集落样生长,黑素细胞的树突较多。培养基1(M1)中贴壁细胞数目少,死亡细胞多,细胞增殖缓慢,角质形成细胞散在,黑素细胞多是双极。培养基3(M3)细胞生长情况介于M1和M2两组之间。结论培养基中的高水平ET-1有利于表皮细胞的贴壁、增殖和分化。在培养过程中表皮细胞能够分泌ET-1,并释放进入培养上清,表皮细胞培养消耗SCF,但无明显分泌作用。
[Abstract]:Objective to observe the growth of epidermal cells in three media and to detect the levels of endothelin-1 (ET-1) and stem cell factor (SCF) at different time points. Methods Epidermal cells were cultured in three kinds of culture medium. The supernatants were collected at 24 h, 48 h, 72 h and 96 h, respectively. The levels of ET-1 and SCF were detected by enzyme-linked immunosorbent assay (ELISA), and the growth status of the cells in different media was compared. Results the content of ET-1 in culture supernatant of culture medium 2 (M2) was the highest in the three groups, and the content of ET-1 increased with time (P0.01), and the content of SCF in the supernatant of culture medium 2 (M2) decreased significantly at 48h after 24 h of culture, and the content of SCF in the supernatant of culture medium 2 (M2) increased with the increase of time (P0.01). The differences among the three groups were statistically significant (P 0.01). The changes of ET-1 and SCF were basically the same before the change of solution (P0.01). The proliferation of M2 cells was the fastest, keratinocytes grew like colony, and melanocytes had more dendrites. In culture medium 1 (M1), the number of adherent cells was small, the number of dead cells was more, the proliferation of cells was slow, the keratinocytes were scattered, and the melanocytes were mostly bipolar. The cell growth of medium 3 (M 3) was between M 1 and M 2 groups. Conclusion the high level of ET-1 in culture medium is beneficial to the adhesion, proliferation and differentiation of epidermal cells. Epidermal cells can secrete ET-1, and release into the supernatant of culture. Epidermal cell culture consumes SCF, but has no effect on secretion.
【作者单位】：苏州大学附属第三医院皮肤性病科;蚌埠医学院第一附属医院皮肤性病科;
【基金】：国家自然科学基金资助项目(81171516)
【分类号】：R751

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