大鼠触须毛乳头细胞的分离和培养

Isolation and in vitro culture of follicular papilla cells from rat vibrissae

目的寻求一种简单有效的分离培养大鼠触须毛乳头细胞的方法。方法将大鼠触须毛囊完整拔出后,在37℃缓慢搅拌的条件下加入0.2%胶原酶Ⅰ消化3h,间断吹打使毛乳头游离,收集上清低速离心(300r/min)后收集毛乳头进行培养,计算其贴壁率,检测α-平滑肌肌动蛋白表达。结果全毛囊消化法能显著降低工作强度,减少污染机会,具有毛乳头贴壁快和细胞迁出时间短的优点。α-平滑肌肌动蛋白染色阳性。结论全毛囊消化法是一种简单有效的分离培养大鼠触须毛乳头细胞的方法。

Objective To establish a simple method for isolating and culturing follicular papilla cells from rat vibrissae. Methods The intact follicles were obtained and digested in 0.2% collagenase I with agitation on a rotary stirrer at 100 r/min at 37 ℃ for 3 h. The suspension was centrifuged at 300 r/min and the papilla cells were collected and suspended in DMEM for cell culture. The adhesion efficiency of the dermal papilla cells was evaluated and compared with that of the cells obtained by microdissection. Results and Conclusion The described procedure allowed efficient and rapid isolation of the dermal papilla cells from rat vibrissae and ensured improved adhesion of the dermal papillae and outgrowth of the cells with reduced labor and risk of contamination. The cells obtained with this procedure were positive for alpha-smooth muscle actin staining.