A simplified method for isolating murine primary pancreatic acinar cells
10.3760/cma.j.issn.1674-1935.2018.02.012
- VernacularTitle:小鼠原代胰腺腺泡细胞分离提纯的简化方法
- Author:
Lu ZHUANG
1
,
2
;
Jianhua WAN
;
Haojie HUANG
;
Zhaoshen LI
;
Baoan JI
Author Information
1. 200433 上海,第二军医大学长海医院消化内科
2. 美国梅奥医学中心佛罗里达分部肿瘤生物学系
- Keywords:
Pancreas;
Acinar cells;
Primary cell;
Cell separation;
Mice
- From:
Chinese Journal of Pancreatology
2018;18(2):112-115
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore a simplified and economical method to isolate the murine primary pancreatic acinar cells.Methods The collagenase and trypsin inhibitor dissolving in DMEM solution were used to digest the murine pancreas,and 4％ BSA dissolving in DMEM solution was used to purify and isolate primary pancreatic acinar cells from pancreas.CCK-8 method was applied to check the ability of pancreatic acinar cells to secret amylase.Results After digestion,shaking in the water bath,resuspension,filtration and precipitation,murine primary pancreatic acinar cells could be obtained within 2 hours.Pancreatic acinar cells in good conditions appeared in clusters,and their basolateral domains were round and devoid of blebs,and the cytoplasm appeared clear.Their apical domain were surrounded by hundreds of zymogen granules which looked darker.The nucleus was located in the basal area of the vesicular region.The basal level of amylase release as a percent of total release from pancreatic acinar cells was around 2.5％ in CCK8-unstimulated group.This rate started to increase after CCK-8 stimulation and reached its peak [(12.83 ± 1.04) ％] at a concentration of 50 pmol/L of CCK-8,but the ratio of the amylase level secreted by the pancreatic acinar cells to the total amylase level displayed a decreasing trend with the increase of CCK-8 concentration.Conclusions This optimized method had the advantage of being fast and simple,low technical difficulty and good repetition.It was a new simplified and cheap method for isolating murine pancreatic acinar cells.
