摘要
目的 :探索肌腱组织块培养法中腱细胞的生长规则及形态 ,为腱细胞培养提供一种简便、可靠的方法。方法 :取家兔 2 0只 ,体重 1.5～ 2 .5kg ,应用显微外科技术剥除屈肌腱外膜组织后 ,以胰酶及胶原酶消化去除残存腱外膜组织 ,腱组织剪成 1mm3 小块 ,放入 5 0ml培养瓶中 ,向瓶内加入少量培养液 (约 1ml) ,培养液由F -12培养基加 2 0 %胎牛血清、青霉素 6mg/L、链霉素 0 .1g/L及抗坏血酸 30mg/L组成。用无菌探针将组织块均匀分置 ,小心置入 37℃、5 %CO2 培养箱中培养 ,2 4h后组织块与瓶壁粘着后再加入F - 12完全培养液。当融合成单层细胞后进行传代培养 ,并以 10 0 0转 /min速度离心淘洗 10min ,去除上层液体 ,向沉淀物加入F -12完全培养液 ,吸管吹打后计数吸取 ,按 1∶2分瓶传代培养。结果 :组织块培养法成功率 95 % ,腱细胞自组织块内爬出贴壁生长时间平均为 (12± 1.7)d左右 ,细胞倍增时间平均为 (6± 0 .9)d。结论 :组织块方法兔腱细胞培养简便、易行。
Objective:To explore the growth and morphology of the tenocytes,and provide an easy,dependable culture method. Method:Twenty rabbits(wt, 1.5～2.5kg)were operated;the peritenon of flexor tendon were removed by microsurgical technique;the tendon were cut into 1mm 3 and then put into a 50 ml culture bottle,added with 1ml culture medium containing F-12 medium,20% FBS,6mg/L penicilin, 0.1g/L streptomycin and 30mg/L ascorbic,then cultured at the condition of 37℃ and with 5% CO; F-12 total culture medium was added 24 hours later. When the cells became monolayer, we subcultured them and counted the cell number. Result: The rate of success of tissue culture method was 95%. The time of tenocytes beginning to grow adhering to the wall was 12±1.7 days; the time of cellular group duplication was 6±0.9 days.Conclusions: Using tissue culture method to culture rabbit tenocytes is an easy way.
出处
《中国现代医学杂志》
CAS
CSCD
2003年第11期39-41,共3页
China Journal of Modern Medicine
