Pancreatic Research Group, South Western Sydney Clinical School and School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia.
Pancreatology. 2010;10(4):434-43. doi: 10.1159/000260900. Epub 2010 Aug 20.
Pancreatic stellate cells (PSCs) play a critical role in pancreatic fibrosis. To date, human PSC biology has been studied using cancer- or inflammation-associated (pre-activated) PSCs, but an in vitro model of quiescent normal human PSCs (NhPSCs) has been lacking.
To (i) isolate and characterize quiescent NhPSCs, and (ii) evaluate the response of culture-activated NhPSCs to cytokines and LPS.
Quiescent NhPSCs were isolated from normal pancreatic tissue using density gradient centrifugation. PSC markers, glial fibrillary acidic protein (GFAP), desmin, α-smooth muscle actin (αSMA) and the lipopolysaccharide (LPS) receptors TLR4 and CD14 were identified by immunoblotting and immunocytochemistry. The effect of platelet-derived growth factor (PDGF), transforming growth factor β (TGFβ) and LPS on NhPSC activation was also assessed.
Freshly isolated NhPSCs displayed a polygonal appearance with refringent cytoplasmic lipid droplets. Culture-activated NhPSCs expressed GFAP, desmin, αSMA, TLR4 and CD14, and were responsive to PDGF, TGFβ and LPS.
Isolated NhPSCs expressed the same markers as rat PSCs and human cancer-associated PSCs and responded to PDGF and TGFβ similarly to rat PSCs. NhPSC preparations provide a useful in vitro tool to study the biology of PSCs in their physiological, non-activated state. and IAP.
胰腺星状细胞（PSCs）在胰腺纤维化中起着关键作用。迄今为止，人们使用与癌症或炎症相关的（预激活的）PSC 来研究人类 PSC 生物学，但缺乏静息正常人类 PSCs（NhPSCs）的体外模型。
（i）分离和鉴定静息 NhPSCs，（ii）评估培养激活的 NhPSCs 对细胞因子和 LPS 的反应。
使用密度梯度离心法从正常胰腺组织中分离静息 NhPSCs。通过免疫印迹和免疫细胞化学鉴定 PSC 标志物胶质纤维酸性蛋白（GFAP）、结蛋白、α-平滑肌肌动蛋白（αSMA）和脂多糖（LPS）受体 TLR4 和 CD14。还评估了血小板衍生生长因子（PDGF）、转化生长因子β（TGFβ）和 LPS 对 NhPSC 激活的影响。
新分离的 NhPSCs 呈多角形外观，细胞质脂滴折光。培养激活的 NhPSCs 表达 GFAP、结蛋白、αSMA、TLR4 和 CD14，并对 PDGF、TGFβ 和 LPS 有反应。
分离的 NhPSCs 表达与大鼠 PSCs 和人类癌症相关 PSCs 相同的标志物，并对 PDGF 和 TGFβ 的反应与大鼠 PSCs 相似。NhPSC 制剂为研究 PSCs 在生理非激活状态下的生物学特性提供了有用的体外工具。
