差速贴壁法分离人子宫腺肌病病灶细胞的平滑肌细胞的纯化时间探讨  

Exploration on the Purification Time of Getting Smooth Muscle Cells from Human Adenomyosis Focus Cells by Different-Speed Adhesion Method

作　　者：许春燕 范为之 宋阳[2] 关永格 李坤寅 XU Chunyan;FAN Weizhi;SONG Yang;GUAN Yongge;LI Kunyin(Guangdong Hospital of Traditional Chinese Medicine,Guangzhou 510120,China;College of Nursing,Guangzhou University of Traditional Chinese Medicine,Guangzhou 510006,China;The Third Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510378,China)

机构地区：[1]广东省中医院,广东广州510120 [2]广州中医药大学护理学院,广东广州510006 [3]广州中医药大学第三附属医院,广东广州510378

出　　处：《临床医学工程》2020年第7期869-870,共2页Clinical Medicine & Engineering

摘　　要：目的探讨差速贴壁法获取人子宫腺肌病病灶细胞的平滑肌细胞的纯化时间。方法采取组织块酶消化法进行子宫腺肌病病灶细胞原代培养,待生长融合80%~90%时根据细胞贴壁时间分为未处理组及15 min、 30 min、 60 min、 90 min四个时间组,各时间贴壁组与未贴壁组进行细胞筛选与培养,以免疫组化方法鉴定平滑肌细胞及其纯度。结果 400倍镜视野下每组16个视野计数算滑肌细胞数百分比,未处理组最低,为12.60%;未贴壁15 min组最高,为73.38%,与未处理组比较有统计学差异(P <0.01);贴壁90 min组与未处理组比较无统计学差异(P>0.05)。结论用差速贴壁法分离人子宫腺肌病病灶的平滑肌细胞可通过贴壁15 min剔除大部分杂质细胞,选择15 min内未贴壁细胞继续培养传代可进一步获取纯度更高的子宫平滑肌细胞。Objective To explore the purification time of getting smooth muscle cell from human denomyosis focus cells by different-speed adhesion method.Methods The human adenomyosis focus cells were primarily cultured by collagenase.When the cells proliferated to 80%~90%,they were divide into untreated group and four time groups(15 min,30 min,60 min and 90 min)according to differential adhesion time.All groups were screened and cultured.Immunohistochemistry was used to identify the smooth muscle cells and the purity.Results Under 400×mirror,the percentage of smooth muscle cells was counted from 16 views of each group.The untreated group was lowest(12.60%);The non-adhesion 15 min group was highest(73.38%),which had statistical difference with the untreated group(P<0.01);No statistical difference wad found between adhesion 90 min group and untreated group(P>0.05).Conclusions Different-speed adhesion method to separate the smooth muscle cell from human denomyosis focus cells can remove most of the impurity cells by 15 min adhesion.More pure smooth muscle cells can be obtained by further culture and subculture of non-adhesion cells within 15 min.

关 键 词：差速贴壁法 子宫腺肌病病灶 平滑肌细胞 纯化 

分 类 号：R711.74[医药卫生—妇产科学]