Modified tissue culture for C57BL/6 mouse pancreatic stellate cells

Objective To culture C57BL/6 mouse pancreatic stellate cells (PSCs) with a modified method, and to establish a simple cultivation method with a high yield of PSCs. Methods Pancreatic tissues in C57BL/6 mice were collected, washed by fetal bovine serum, minced into 0.5~1 mm3, and cultured in sterile culture flasks. After 4days, primary PSCs were collected and passed on. The changes in intracellular lipid droplets were identified by oil red O staining; α-SMA, desmin expression was detected by immunocytochemical or immunocytofluorescent staining, and Western blot. Results Outgrowth cells in the fourth day were oval or star with abundant lipid droplets and fewer cells expressed α-SMA and desmin protein. PSCs turned into bigger, more pseudopodial ones with decreasing lipid droplets and significantly increasing expression of α-SMA and desmin protein after the passage. Protein expression of α-SMA in primary and passage PSCs were 0.653±0.071, 2.290±0.055, respectively; and the expression in passage PSCs was significantly higher than that in primary PSCs (P<0.05), and the number of cells expressing desmin protein was significantly increased. Conclusions Modified tissue culture is a high yield and high purity method for both quiescent and activated PSCs, which can meet the need of in-vitro studies. Key words: Tissue cutture techniques; Pancreatic stellate cells; Cell separation; In vitro