BACKGROUND: 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. AIMS: To (i) isolate and characterize quiescent NhPSCs, and (ii) evaluate the response of culture-activated NhPSCs to cytokines and LPS. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
BACKGROUND: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. AIMS: To (i) isolate and characterize quiescent NhPSCs, and (ii) evaluate the response of culture-activated NhPSCs to cytokines and LPS. METHODS: 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. RESULTS: 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. CONCLUSION: Isolated NhPSCs expressed the same markers as rat PSCs and humancancer-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.
Authors: Shadan Ali; Raagini Suresh; Sanjeev Banerjee; Bin Bao; Zhihong Xu; Jeremy Wilson; Philip A Philip; Minoti Apte; Fazlul H Sarkar Journal: Am J Cancer Res Date: 2015-02-15 Impact factor: 6.166
Authors: Elizabeth R Murray; Shinelle Menezes; Jack C Henry; Josie L Williams; Lorena Alba-Castellón; Priththivika Baskaran; Ivan Quétier; Ami Desai; Jacqueline J T Marshall; Ian Rosewell; Marianthi Tatari; Vinothini Rajeeve; Faraz Khan; Jun Wang; Panoraia Kotantaki; Eleanor J Tyler; Namrata Singh; Claire S Reader; Edward P Carter; Kairbaan Hodivala-Dilke; Richard P Grose; Hemant M Kocher; Nuria Gavara; Oliver Pearce; Pedro Cutillas; John F Marshall; Angus J M Cameron Journal: Cell Rep Date: 2022-01-25 Impact factor: 9.423