目的探讨用微载体培养技术进行人毛乳头细胞高效扩增的培养方法,以满足对毛乳头细胞的大量需求。方法采用Cytodex-3为培养载体进行人第3代毛乳头细胞的微载体培养,定期观察培养液颜色的变化,采用苔盼蓝染色进行活细胞计数,倒置显微镜下观察生长状况。将微载体上消化的细胞进行传统培养,观察细胞生长情况,甲苯胺蓝染色,细胞免疫组织化学染色。结果培养至第12天,微载体上细胞数量最多,并且细胞聚集成团,类似毛乳头样结构。利用这时的细胞进行普通培养,仍然表现出凝集性生长特点,细胞呈异染性,免疫组织化学染色阳性。结论利用Cytodex-3培养可以收获大量具有生物活性的毛乳头细胞。
Objective To establish a more effective method in order to obtain human dermal papilla cells (DPCs) for the need of a large number of DPCs. Method Third-passage of human DPCs were cultured on cytodex-3 microcarriers. The color change of the medium was observed and the cultured cells counted. The growth of DPCs was observed under the inverted microscope. Cells seperated from cytodex-3 were cultured in a traditional way. These cells were stained immunohistochemically or by toluidine blue. Results The density of cultured DPCs was the largest at the 12th day. The cells were positive to smooth muscle actin alpha and prensented metachromatism by tuluidine blue staining. Conclusions DPCs that remain the primary biological feature are obtained densely on cytodex-3.