A modified method for isolating human quiescent pancreatic stellate cells

Abstract

Background: This study explored a simple, high-yield method for isolating quiescent human pancreatic stellate cells (PSCs) to provide sufficient and reliable raw materials for PSC-related studies. Materials and methods: Single-cell suspensions were prepared from normal human pancreatic tissue specimens using the gentleMACS ™ tissue processor, which enhanced the yield and viability of the suspensions. Percoll density gradient centrifugation was then performed to isolate quiescent normal PSCs (NPSCs). Cell viability was determined by trypan blue staining, and the states of the NPSCs were determined by autofluorescence and oil red O staining. The purity of human activated PSCs (APSCs) was determined by immunofluorescence assays. Results: The yield of NPSCs was ~(2.75±0.65)×10 6 cells/g. The maximum cell viability was 92%, whereas the maximum cell purity was 95%. Conclusion: The method employed in this study to isolate PSCs is a simple, high-yield and stable method that is worth popularizing.