A new method for separation of human pancreatic stellate cells

Abstract

Objective To explore a new method for the separation of human pancreatic stellate cells. Methods Single-cell suspension of normal pancreatic tissue and pancreatic cancer tissue was prepared by gentle MACS™ tissue processor-constant temperature shaking digestion. Human pancreatic stellate cells of quiescent and activated state were isolated by density gradient centrifugation. Results A new type of isolation method could obtain about (2.6±0.7) ×106 quiescent pancreatic stellate cells in 1 g of human normal pancreatic tissue, with a viability of about 90.0%. The morphology of the cells were conformed to the representative for the quiescent state characteristics and transient blue-green autofluorescence was observed at the 328 nm excitation wavelength; 1 g of human pancreatic cancer was able to obtain approximately (4.1±1.1)×106 activated PSCs with a viability of 92.0%, and all of the activated cells expressed α-SMA vimentin, FSP-1 and other characteristic markers. Conclusions The new separation method of this experiment is suitable for both human resting and activated human pancreatic stellate cells. At the same time, the purity is high and the separation time is greatly shortened, which is worth promoting. Key words: Pancreatic stellate cells, human; Separation method; Technological improvement; Density gradient centrifugation