组织工程皮肤中人毛乳头细胞体外培养条件的优化

Optimization of the culture in vitro of human dermal papilla cells in the tissue-engineered skin

目的:优化人毛乳头细胞体外培养条件。方法:采用成人头皮标本,显微器械分离法及器械分离结合胶原酶消化二步法分离毛乳头,分别利用普通有血清的Dulbecco最低必需培养基(DMEM)、含生长因子的有血清DMEM培养基及角质细胞条件培养基,进行人毛乳头细胞的体外培养,比较细胞的生长特点、增殖活性、细胞周期等生物学特性。结果:二步法分离毛乳头的效率为显微器械分离法的100倍,利用含生长因子的有血清DMEM培养基和角质细胞条件培养基均可使毛乳头细胞长期维持较强的增殖潜能。结论:二步法分离毛乳头,利用含生长因子的有血清DMEM培养基或角质细胞条件培养基进行培养,可以快速、简便、高效地获得大量保持高增殖潜能的毛乳头细胞。

AIM:To optimize the culture of human dermal papilla cells in vitro. METHODS:Dermal papillas were isolated from specimens of adults' scalp with the method of micro dissection or the method of dissection followed by collagenase digestion,then the dermal papilla cells were cultured in vitro with Dulbecco's modified eagle medium(DMEM) containing 100 mL/L serum,DMEM containing 100 mL/L serum and growth factors,or keratinocyte conditioned medium respectively.The biological characteristics of the above cells were compared in growth behavior,proliferative activity and cell cycle. RESULTS:The efficiency of isolating dermal papilla with the method of dissection followed by collagenase digestion was higher than the method of micro dissection by 100 times.The dermal papilla cells showed high or proliferative activity lasting for a long term in the DMEM containing serum and growth factors and the keratinocyte conditioned medium. CONCLUSION:Isolated by means of dissection followed by collagenase digestion,and then cultured in the DMEM containing serum and growth factors or the keratinocyte conditioned medium,a mass of dermal papilla cells with high proliferative potential can be obtained promptly,simply and efficiently.