Tissue explant methodversus enzymatic digestion method for culture of rat hair follicle stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Hair folicle stem cels have been confirmed to have stronger proliferative ability than interfolicular epidermal stem cels, which have been an issue of concern in seed cel research. OBJECTIVE:To compare the biological characteristics of rat hair folicle stem cels cultured by tissue explant method and enzymatic digestion method. METHODS: Under stereomicroscope, hair folicles were isolated from the rat whiskers, and then tissue explant method and two-step enzymatic digestion method were employed to culture hair folicle stem cels. Cels were purified using repeated differential adhesion method, and cel growth and morphology were observed periodicaly. Flow cytometry was used to detect the expression of CD34 and β1 integrin in passage 3 hair folicle stem cels. RESULTS AND CONCLUSION:Cels cultured by two-step enzymatic digestion method grew faster with more amount than those cultured by tissue explants method. Flow cytometry showed that the expressions of PE-CD34 and FITC-β1 were (39.52±19.57)% and (93.46±4.73)% for the two-step enzymatic digestion group, and (19.20±11.53)% and (363.57±14.42)% for the tissue explant method, respectively. There was a significant difference between the two methods. In conclusion, these two methods are able to culture high-activity hair folicle stem cels, which can be chosen according to different experimental requirements.