[1]郑幸龙,刘雯雁,刘锋锋,等.人脂肪肝脱细胞支架的制备及肝癌细胞的体外三维培养[J].南方医科大学学报,2019,(08):930.[doi:10.12122/j.issn.1673-4254.2019.08.09]
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人脂肪肝脱细胞支架的制备及肝癌细胞的体外三维培养()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2019年08期
页码:
930
栏目:
出版日期:
2019-08-31

文章信息/Info

Title:
Decellularized matrix of human fatty liver used for three-dimensional culture of hepatocellular carcinoma cells
作者:
郑幸龙刘雯雁刘锋锋李静向俊西刘鹏吕毅
关键词:
人脂肪肝脱细胞支架三维培养肝癌模型
Keywords:
human fatty liver decellularized matrix three-dimensional culture hepatocarcinoma cell model
DOI:
10.12122/j.issn.1673-4254.2019.08.09
摘要:
目的体外建立一个基于人脂肪肝脱细胞支架的肝癌三维模型。方法通过反复冻融、梯度浓度的SDS和1% Triton X-100 经门静脉和肝动脉双重灌注及组织块在Triton X-100中反复震荡的方法制备人脂肪肝脱细胞基质(hFLM)。在hFLM内培养 HepG2细胞,检测其存活、形态、增殖及黏附分子表达情况。结果采用双重灌注的方式显著缩短了支架制备的时间,同时保留 了肝脏的细胞外基质成分及三维结构。HepG2细胞在hFLM内培养15 d,存活良好,并呈现与体内相似的肿瘤增殖模式。而 且,hFLM培养的HepG2细胞和皮下成瘤组均低表达E-cadherin,高表达Vimentin,与二维培养组正好相反。基于hFLM的肝癌 模型缺少有效的血管网络,缺乏足够的营养转运,可能导致后期细胞增殖减缓。hFLM培养的HepG2细胞第12天时PCNA指数 较第6天时降低了29.3%。结论建立一种新的人脂肪肝脱细胞方案,并验证了基于hFLM体外构建肝癌模型的可行性。
Abstract:
Objective To construct a decellularized matrix of human fatty liver as the scaffold for three-dimensional (3D) culture of hepatocarcinoma cells. Methods Human fatty liver decellularized matrix (hFLM) was prepared by repeated freezingthawing, perfusion with gradient SDS and 1% Triton X-100 through the portal vein and hepatic artery, and repeated agitation with Triton X-100. HepG2 cells were cultured in the prepared hFLM, and the cell survival, morphology, proliferation and cellular expressions of the adhesion molecules were detected. Results The decellularization procedure shortened the time for scaffold preparation and preserved the 3D ultrastructure and the composition of the extracellular matrix. HepG2 cells cultured in hFLM scaffold maintained proliferation for up to 15 days and showed a growth pattern with a long lag phase and a slow growth rate, which was similar to the growth pattern in vivo. The cultured HepG2 exhibited a low expression of E-cadherin and a high expression of vimentin, which was consistent with the xenograft but opposite to 2D cultured cells. However, the lack of adequate nutrient transport in this hepatocarcinoma cell model led to a slowdown of cell proliferation in the later stage. The PCNA index of HepG2 cells cultured in hFLM was lowered by 29.3% on day 12 as compared with that on day 6. Conclusion We established a new protocol for preparing hFLM and confirmed the feasibility of constructing hepatocarcinoma cell models using the hFLM scaffold.
更新日期/Last Update: 1900-01-01