title: Hepatocyte culture in autologous decellularized spleen matrix.
abstract: OBJECTIVE
  Using decellularized scaffold to reengineer liver tissue is a promising alternative therapy for end-stage liver diseases. Though the decellularized human liver matrix is the ideal scaffold for reconstruction of the liver theoretically, the shortage of liver donors is still an obstacle for potential clinical application. Therefore, an appropriate alternative scaffold is needed. In the present study, we used a tissue engineering approach to prepare a rat decellularized spleen matrix (DSM) and evaluate the effectiveness of this DSM for primary rat hepatocytes culture.
  
  
  METHODS
  Rat decellularized spleen matrix (DSM) was prepared by perfusion of a series of detergents through spleen vasculature. DSM was characterized by residual DNA and specific extracellular matrix distribution. Thereafter, primary rat hepatocytes were cultured in the DSM in a 3-dimensional dynamic culture system, and liver cell survival and biological functions were evaluated by comparison with 3-dimensional sandwich culture and also with cultured in decellularized liver matrix (DLM).
  
  
  RESULTS
  Our research found that DSM did not exhibit any cellular components, but preserved the main extracellular matrix and the intact vasculature evaluated by DNA detection, histology, immunohistochemical staining, vessel corrosion cast and upright metallurgical microscope. Moreover, the method of DSM preparation procedure was relatively simple with high success rate (100%). After seeding primary hepatocytes in DSM, the cultured hepatocytes survived inside DSM with albumin synthesis and urea secretion within 10 d. Additionally, hepatocytes in dynamic culture medium had better biological functions at day 10 than that in sandwich culture. Albumin synthesis was 85.67 ± 6.34 μg/10(7) cell/24h in dynamic culture in DSM compared to 62.43 ± 4.59 μg/10(7) cell/24h in sandwich culture (P < 0.01) and to 87.54 ± 5.25 μg/10(7) cell/24h in DLM culture (P > 0.05); urea release was 32.14 ± 8.62 μg/10(7) cell/24h in dynamic culture in DSM compared to 20.47 ± 4.98 μg/10(7) cell/24h in sandwich culture (P < 0.05) and to 37.38 ± 7.29 μg/10(7) cell/24h cultured in DLM (P > 0.05).
  
  
  CONCLUSIONS
  The present study demonstrates that DSM can be prepared successfully using a tissue engineering approach. The DSM is an appropriate scaffold for primary hepatocytes culture.
authors: a Medical College; Xi'an Jiaotong University ; Xi'an , Shaanxi , China.
authors: Rui
authors: R
authors: Gao
authors: Wanquan
authors: W
authors: Wu
authors: Junxi
authors: J
authors: Xiang
authors: Yi
authors: Y
authors: Lv
authors: Xinglong
authors: X
authors: Zheng
authors: Qian
authors: Q
authors: Chen
authors: Haohua
authors: H
authors: Wang
authors: Bo
authors: B
authors: Wang
authors: Zhengwen
authors: Z
authors: Liu
authors: Feng
authors: F
authors: Ma
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry; D003902:Detergents / Q000737:chemistry; D005109:Extracellular Matrix / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000473:pathology; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D013154:Spleen; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D003902:Detergents; D004247:DNA
keywords: decellularized spleen matrix; hepatocytes culuture; liver; spleen; tissue engineering