中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2009年
16期
1135-1138
,共4页
肝脏%生物支架%去细胞化
肝髒%生物支架%去細胞化
간장%생물지가%거세포화
Liver%Biological scaffold%Decellularization
目的 通过去细胞化技术建立保留细胞外基质的完整肝脏支架,并对支架进行形态结构和保留成分鉴定.方法 通过门静脉插管,依次灌注去垢剂曲拉通X-100(Triton X-100),十二烷基硫酸钠(SDS),并用磷酸盐缓冲液(PBS)洗脱残留去垢剂,对所得支架进行HE染色、门静脉及月日道铸型、扫描电镜、纤维成分染色鉴定等形态学观察.并将支架切成50 μm的厚度后与C3A细胞系共培养,进行生物相容性验证.结果 经本实验方案得到肝脏生物支架的成功率为75%.肝脏生物支架包膜完整,肉眼可见肝脏内Gllisson管道结构.HE染色示无细胞及胞核残存.管道铸型见胆道、门静脉完整,无铸型液溢出.扫描电镜示大量纤维网状结构存在.纤维成分染色见大量胶原纤维、弹力纤维存在.生物相容性实验初步显示该支架具备较好的生物相容性,可以作为生物支架材料应用.结论 经本实验去细胞化过程,肝组织细胞可从细胞外基质中完全洗脱下来,并保留比较完整的肝脏管道系统.本研究获得全肝生物支架可以作为肝脏器官培养的基础支架.
目的 通過去細胞化技術建立保留細胞外基質的完整肝髒支架,併對支架進行形態結構和保留成分鑒定.方法 通過門靜脈插管,依次灌註去垢劑麯拉通X-100(Triton X-100),十二烷基硫痠鈉(SDS),併用燐痠鹽緩遲液(PBS)洗脫殘留去垢劑,對所得支架進行HE染色、門靜脈及月日道鑄型、掃描電鏡、纖維成分染色鑒定等形態學觀察.併將支架切成50 μm的厚度後與C3A細胞繫共培養,進行生物相容性驗證.結果 經本實驗方案得到肝髒生物支架的成功率為75%.肝髒生物支架包膜完整,肉眼可見肝髒內Gllisson管道結構.HE染色示無細胞及胞覈殘存.管道鑄型見膽道、門靜脈完整,無鑄型液溢齣.掃描電鏡示大量纖維網狀結構存在.纖維成分染色見大量膠原纖維、彈力纖維存在.生物相容性實驗初步顯示該支架具備較好的生物相容性,可以作為生物支架材料應用.結論 經本實驗去細胞化過程,肝組織細胞可從細胞外基質中完全洗脫下來,併保留比較完整的肝髒管道繫統.本研究穫得全肝生物支架可以作為肝髒器官培養的基礎支架.
목적 통과거세포화기술건립보류세포외기질적완정간장지가,병대지가진행형태결구화보류성분감정.방법 통과문정맥삽관,의차관주거구제곡랍통X-100(Triton X-100),십이완기류산납(SDS),병용린산염완충액(PBS)세탈잔류거구제,대소득지가진행HE염색、문정맥급월일도주형、소묘전경、섬유성분염색감정등형태학관찰.병장지가절성50 μm적후도후여C3A세포계공배양,진행생물상용성험증.결과 경본실험방안득도간장생물지가적성공솔위75%.간장생물지가포막완정,육안가견간장내Gllisson관도결구.HE염색시무세포급포핵잔존.관도주형견담도、문정맥완정,무주형액일출.소묘전경시대량섬유망상결구존재.섬유성분염색견대량효원섬유、탄력섬유존재.생물상용성실험초보현시해지가구비교호적생물상용성,가이작위생물지가재료응용.결론 경본실험거세포화과정,간조직세포가종세포외기질중완전세탈하래,병보류비교완정적간장관도계통.본연구획득전간생물지가가이작위간장기관배양적기출지가.
Objective To explore Sift innovative method for preparing a whole-liver reconstruct scaffold with intact three-dimensional geometry, vasculature and bile duct by decellularization technology.Methods The portal vein was annulated and perfused sequenfially with 1% Triton X-100 and 1% SDS for about 4 h,and then was perfused with phosphate buffered saline to dilute SDS residue.The retained stmcRlre was evaluated by histological analyses,including macroscopic,Hematoxylin-Eosin staining,Masson's trichrorne staining,orcein staining and SEM.The liquid polymer preparation 8%-10%,which was made of chlorinated poly vinyl chlofide(CPVC as solute),acetone(as solvent)and pigment,was injected into portal vein and bile duct to demonstrate tIle integrity of the portal vein and bile duct. The scaffold was cut into slices with the thickness of about 50μm and co-cultured with C3A cell line.Results Macroscopic examination showed that the deedlularized liver WS8 transparent and intrahepatic Gli8son's system could be observed.H&E staining of slices of decellularized liver demonstrated no intact cells or nuclei existect Masson trichrome staining revealed collagen retained.Orcein staining showed that there were elastic fibers.SEM showed the network of ECM Was intacL C3A-to-scaffold co-culture revealed the scaffold of a good biocompatibility.Conclusion Peffusion with detergents throuSh portal vein for liver decellularization is an efllcient method to obtain a completely whole-1iver scaffold for hepatic organ reconstruction.
