新型全肝脏去细胞化流程对细胞外基质及细胞组分残留量影响的研究

doi:10.3969/j.issn.0258-8021.2013.01.009

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Abstract There has been a conflict between preservation of extracellular matrix and reduction of donor’s cell components on decellularized scaffold production in tissue engineering. Therefore a protocol for decellularization is the key to solve this problem. We developed a novel decellularization protocol which used TritonX-100 as a major agent for decellularizing assisted by SDS, enzymes, freezethaw processing, hypotonic and hypertonic solutions etc. After morphological and histological observation through immunohistochemistry and scanning electron microscope, evaluation of ECM content by ELISA, using absorbance detection and Westernblotting to test cell components residuals, and immunofluorescence after 3D perfusion culture of WBF-344 cells in the scaffold, the results showed that, by using this new protocol, three dimensional structure like collagen, laminin, fibronectin appeared well protected, and the extracellular matrix contents of decellularized scaffold such as GAG, HGF and VEGF was 90% and 40% of normal tissue. Besides a DNA elimination, MCHI and HMGB-1 were only 10% of normal tissue. WBF-344 cells grew well in the scaffold and the albumin level was rising. In conclusion, this study built a novel decellularization protocol and evaluation system, and proposed a promising application of the acellular liver scaffold in stem cell differentiation and liver disease therapy.

Key words： decellularized scaffold extracellular matrix cell components residuals perfusion culture stem cell differentiation

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	WANG Yang¹LIU Zan¹YE HaiLin ¹FU JunHui¹XU QiGang^1ZENG QiQiang¹WU JianBo²ZHANG QiYu^1*

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WANG Yang¹LIU Zan¹YE HaiLin ¹FU JunHui¹XU QiGang^1ZENG QiQiang¹WU JianBo²ZHANG QiYu^1*. The Effect of a Novel Whole Liver Decellularization Protocol on the Extracellular Matrix and Cell Components Residuals[J]. journal1, 2013, 32(1): 61-69.

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http://cjbme.csbme.org/EN/10.3969/j.issn.0258-8021.2013.01.009 OR http://cjbme.csbme.org/EN/Y2013/V32/I1/61

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