Influencing factors on the primary cultur e of mouse fibroblast-like synov iocyt es
- VernacularTitle:小鼠成纤维样滑膜细胞原代培养的影响因素研究
- Author:
Mei TANG
;
Shan CHANG
;
Zongrui CAO
;
Xingyan LUO
;
Song HU
;
Xinwei TANG
;
Yantang WANG
;
Yang LIU
;
Qiang ZOU
- Publication Type:Journal Article
- Keywords:
Fibroblast-like synoviocyte;
Culture invi tro;
Cell identification;
Mouse
- From:
Journal of Medical Postgraduates
2014;(8):789-792
- CountryChina
- Language:Chinese
-
Abstract:
Obej ctive Knockout mice are widely used in the studies of joint diseases .This article investigated the effects of joint processing methods , collagenase types ,and collagenase digestion time on the number of primary fibroblast -like synoviocytes (FLSs) obtained from mice. Methods The hind legs of 6 of the 12 male mice were cut open from the hip joints , but not those of the other 6.FLSs were isolated using the type-Ⅳcollagenase digestion method and purified by differential digestion .Cell morphology was observed under the inverted microscope .The type, viability, and purity of the cells were determined by flow cytometry . Rse ults Significantly fewer FLSs were obtained from the mice with the hind legs cut open ( 19 133 ±115 ) than from those without (24 933 ± 503) (P<0.05).The numbers of FLSs collected from the cell suspension at 1, 2, 3, 4, 5,6 , and 7 hours after digestion were 700 ±300 , 600 ±100 , 15 200 ±900 , 5100 ±800 ,2700 ±300 , 900 ±200, and 300 ±100, respectively, the highest at 3 hours. There were statistically significant differences in the total number of FLSs obtained by type-Ⅳ and type-Ⅱ collagenase digestions (24900 ±500v s 18 100 ±400, P<0.05). Conclusion For in virt o culture of primary mouse FLSs, it is recommended that the hip joints be not cut open, and type-Ⅳcollagenase be used with cell sus-pension at 2-6hours after digestion .
