血清浓度对皮肤源性前体细胞在生物材料上增殖生长的影响

Effect of media with different serum concentration on proliferation and growth of skin-derived precursors cultured on biomaterials

目的:探讨血清浓度对皮肤源性前体细胞(skin-derived precursors,SKPs)在蝶翅及壳聚糖膜等生物材料表面增殖生长的影响。方法:采用酶消化法从新生大鼠背部皮肤分离纯化SKPs,将传代后的SKPs分别与具有不同表面结构的蝶翅和壳聚糖膜材料共培养,各组材料分别采用含有1%、5%、10%胎牛血清(fetal bovine serum,FBS)浓度的培养基条件,培养4、24、48 h后以细胞活性检测试剂盒(cell counting kit-8,CCK-8)检测各组材料上细胞活力,培养48 h采用倒置显微镜观察SKPs在材料上的贴壁和生长情况。结果:分离纯化的SKPs在各组材料上均能贴壁生长。在蝶翅材料上培养4、24 h后,CCK-8检测结果显示在含有1%FBS浓度培养基条件下SKPs的活力显著低于5%和10%FBS,培养48 h后倒置显微镜下观察,可见1%FBS条件下,细胞大量裂解出现碎片,5%和10%FBS培养条件下,SKPs均生长形态正常,两种蝶翅材料间差异无统计学意义。与蝶翅材料相似,SKPs在具有不同表面形貌的壳聚糖膜各组材料上培养,在1%FBS条件下各种表面形貌壳聚糖膜上SKPs的活力均显著低于5%和10%FBS,且培养48 h,细胞出现大量裂解碎片,5%和10%FBS下SKPs生长形态正常。结论:在具有表面微纳拓扑结构的生物材料上,1%低血清浓度不利于SKPs的生长,5%较低血清浓度可以满足SKPs细胞增殖生长的需要。

Objective: To investigate the effects of serum concentrations on the proliferation and growth of skin-derived precursors(SKPs) on the surfaces of butterfly wings and chitosan membranes. Methods: SKPs isolated and purified from the dorsal skin of newborn rats were co-cultured with different surface morphology materials of butterfly wing and chitosan membrane,respectively. Three serum concentrations of 1%, 5% and 10% fetal bovine serum(FBS) were used for each material. After cultured for 4, 24 and 48 h, the cell proliferation was detected by cell counting kit-8(CCK-8) test. The adhesion and growth of SKPs were observed by inverted microscope after cultured on the biomaterials for 48 h. Results: The SKPs obtained by isolation and purification could adhere and grow on each biomaterial group. After 4 and 24 hours of culture on the butterfly wing material,CCK-8 test results showed that the activity of SKPs under the condition of 1%FBS medium was significantly lower than that of 5% and 10%FBS. After 48 h of culture, large amounts of cell debris were found under the condition of 1%FBS medium by inverted microscope. The growth morphology of SKPs was normal under 5% and 10%FBS culture conditions, and there was no significant difference between the two butterfly wing materials. Similar to the cells on butterfly wings, SKPs were cultured on chitosan membrane materials with different surface morphologies. Under the condition of 1% FBS medium, the activities of SKPs on chitosan membrane with various surface morphologies were significantly lower than those under the conditions of 5%and 10%FBS, and a large number of cell fragmentation appeared after 48 h of culture. SKPs grew well under 5% and 10%FBS culture conditions on all chitosan membranes. Conclusion: The medium with 5% and 10% serum concentration, rather than 1% serum, was suitable for the proliferation and growth of SKPs on biomaterials.