title: Inhibition of pancreatic stellate cell activity by adipose-derived stem cells.
abstract: BACKGROUND
  Pancreatic stellate cells (PSCs) play a critical role in the development of pancreatic fibrosis. In this study we used a novel method to isolate and culture rat PSCs and then investigated the inhibitory effects of adipose-derived stem cells (ADSCs) on activation and proliferation of PSCs.
  
  
  METHODS
  Pancreatic tissue was obtained from Sprague-Dawley rats for PSCs isolation. Transwell cell cultures were adopted for co-culture of ADSCs and PSCs. PSCs proliferation and apoptosis were determined using CCK-8 and flow cytometry, respectively. alpha-SMA expressions were analyzed using Western blotting. The levels of cytokines [nerve growth factor (NGF), interleukin-10 (IL-10) and transforming growth factor-beta1 (TGF-beta1)] in conditioned medium were detected by ELISA. Gene expression (MMP-2, MMP-9 and TIMP-1) was analyzed using qRT-PCR.
  
  
  RESULTS
  This method produced 17.6+/-6.5X10(3) cells per gram of the body weight with a purity of 90%-95% and a viability of 92%-97%. Co-culture of PSCs with ADSCs significantly inhibited PSCs proliferation and induced PSCs apoptosis. Moreover, alpha-SMA expression was significantly reduced in PSCs+ADSCs compared with that in PSC-only cultures, while expression of fibrinolytic proteins (e.g., MMP-2 and MMP-9) was up-regulated and anti-fibrinolytic protein (TIMP-1) was down-regulated. In addition, NGF expression was up-regulated, but IL-10 and TGF-beta1 expressions were down-regulated in the co-culture conditioned medium compared with those in the PSC-only culture medium.
  
  
  CONCLUSIONS
  This study provided an easy and reliable technique to isolate PSCs. The data demonstrated the inhibitory effects of ADSCs on the activation and proliferation of PSCs in vitro.
authors: Department of Hepatobiliary and Pancreatic Surgery, First Affiliated Hospital, Wenzhou Medical College, Wenzhou 325000, China. tangyinhe@sina.com.
authors: Fu-Xiang
authors: FX
authors: Yu
authors: Long-Feng
authors: LF
authors: Su
authors: Chun-Lei
authors: CL
authors: Dai
authors: Yang
authors: Y
authors: Wang
authors: Yin-Yan
authors: YY
authors: Teng
authors: Jun-Hui
authors: JH
authors: Fu
authors: Qi-Yu
authors: QY
authors: Zhang
authors: Yin-He
authors: YH
authors: Tang
mesh_terms: D000199:Actins / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D017209:Apoptosis; D049109:Cell Proliferation; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D015536:Down-Regulation / Q000235:genetics; D015870:Gene Expression; D016753:Interleukin-10 / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000235:genetics; D020780:Matrix Metalloproteinase 9 / Q000235:genetics; D020932:Nerve Growth Factor / Q000378:metabolism; D058954:Pancreatic Stellate Cells / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D053773:Transforming Growth Factor beta1 / Q000378:metabolism; D015854:Up-Regulation / Q000235:genetics
chemical_list: D000199:Actins; D017077:Culture Media, Conditioned; D016753:Interleukin-10; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D020932:Nerve Growth Factor; D019715:Tissue Inhibitor of Metalloproteinase-1; D053773:Transforming Growth Factor beta1; C576839:TIMP1 protein, rat; C498862:Acta2 protein, rat