Isolating, culturing and qualification of mouse pancreatic stellate cells
FAN Xin1, WANG Ying2, HOU Wen-ji3, LIU Yu3, WANG Xu-qing1, SUN Kang1, ZHANG Jian-xin1
(1.Department of General Surgery, 2.Department of Endocrinology, the Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001; 3.School of Clinical Medicine, Jiangsu University, Zhenjiang Jiangsu 212001, China)
Abstract:Objective: To establish a new method to isolate and culture mouse primary pancreatic stellate cells(PSCs). Methods: Based on techniques of isolating and culturing rat PSCs, we acquired purified PSCs from pancreatic tissue fragments via isolation, culture and passage. Oil red O staining, immunofluorescence as well as western blotting were performed to identify pancreatic stellate cells. Cell growth and vitality were validated by cell counting and trypan blue staining. Results: Primary PSCs were positive for oil red O staining; cells grew rapidly and began being activated from the third day. After passage, most of cells were activated. Immunofluorescence staining showed that α-smooth muscle actin (α-SMA) was positive expressed in activated PSCs. Western blotting showed that α-SMA and desmin were expressed in activated PSCs, and that the expression of desmin in fully activated PSCs was obviously higher(P<0.05).Conclusion: This method can be successfully used to isolate mouse PSCs. The yield, purity and activity of PSCs were higher, and this new method could thus meet the research of pancreatic stellate cells in vitro.
范昕,王莹,侯雯跻,等. 小鼠胰腺星状细胞的分离培养与鉴定[J]. 江苏大学学报:医学版, 2012, 22(6): 468-471.
FAN Xin1, WANG Ying2, HOU Wen-ji3, LIU Yu3, WANG Xu-qing1, SUN Kang1, ZHANG Jian-xin1. Isolating, culturing and qualification of mouse pancreatic stellate cells. Journal of Jiangsu University(Medicine Edition), 2012, 22(6): 468-471.
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