ISOLATION AND CULTURE METHOD OF MATURE ADIPOCYTE AND PREADIPOCYTE

摘要

A method for isolating and culturing mature adipocyte and preadipocyte is provided to offer high recovery rate of the adipocyte and preadipocyte by treating fat tissue with a solution composition for each step to isolate the mature adipocyte and preadipocyte in maximum-activated state, and then culturing each of the isolated adipocyte. The method comprises the steps of: (a) washing fat tissue obtained from human body at least three times with 1X-10X phosphate-buffered solution(PBS) where 0.01-1% of bovine serum albumine(BSA) is added; (b) after adding 0.1-0.3% of collagenase type-II solution to the washed fat tissue, agitating it at a temperature of 36-38 deg.C for 10-60 minutes to perform enzymatic treatment; (c) adding Dulbecco's minimum essential medium solution(DMEM), where 1-20% of fetal bovine serum(FBS) is added, to the fat tissue of the step(b) to stop the enzymatic reaction; (d) after adding a fat cell culture solution to the fat tissue of the step(c), centrifuging it to separate the fat tissue into mature adipocyte and preadipocyte; and (e) after respectively collecting the separated mature adipocyte and preadipocyte, respectively culturing it.