| 宾文婷 常加松 朱汉卿 史小林 王小红.胚胎肝细胞用于肝组织工程的体外培养研究[J].,2016,16(19):3629-3633 |
| 胚胎肝细胞用于肝组织工程的体外培养研究 |
| Study on in vitro Culture of Embryonic Hepatocyte for Hepatic TissueEngineering |
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| DOI： |
| 中文关键词: 肝组织工程 胚胎肝细胞 三维立体培养 小鼠 |
| 英文关键词: Hepatic tissue engineering Embryonic hepatocyte 3D culture Mouse |
| 基金项目:江苏省教育厅资助项目（13KJD310001）；南京中医药大学校青年科研课题项目（11XZR02） |
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| 中文摘要: |
| 目的：观察三维受控组装系统下，胚胎肝细胞在三维立体结构的体外生长状态，探讨胚胎肝细胞在肝组织工程中应用的可
行性。方法：用清华大学机械工程系研制的"三维受控组装系统"，将第15 d小鼠胚胎肝细胞作为肝组织工程的种子细胞，与以明
胶为主的复合材料混合，构建成复杂三维立体结构，观察其体外生长发育状态。对体外培养1周及4 周的三维类肝组织标本进行
苏木精- 伊红（HE）染色，免疫组织化学方法检测甲胎蛋白（AFP）及白蛋白（ALB）的表达，并对体外培养4 周的三维类肝组织用
PAS 显色法检测肝糖原表达。结果：HE 染色结果显示体外培养的胚胎肝细胞在三维支架材料中，可形成含有类血管和肝组织样
结构；体外培养1 周的类肝组织AFP 表达呈阳性，体外培养4 周的三维类肝组织ALB 表达呈阳性，PAS显色亦呈阳性。结论：在
三维受控组装系统的构建下，呈立体状生长的胚胎肝细胞，可逐渐形成肝组织样结构，并显示一定的肝脏功能。 |
| 英文摘要: |
| Objective:To observe using embryonic hepatocytes as the seed cells, the growth state of the 3D structure in vitro
which structured by "three dimension (3D) controlled assembly system ", and to investigate the feasibility of embryonic hepatocytes for
hepatic tissue engineering.Methods:We used "3D controlled assembly system" which was developed by Department of Mechanical Engineering
Tsinghua University. We mixed embryonic 15 day mice embryonic hepatocytes with gelatin-based composite materials to assemble
into complex 3D structures. The 3D liver-like tissues were cultured in vitro, and their growing situation were observed by Hematoxylin
and Eosin (HE) staining after cultured for 1 week and 4 weeks. The expression of alpha fetoprotein (AFP) and albumin(ALB)
were evaluated by immunohistochemistry in 3D liver-like tissue section specimens after cultured for 1 week and 4 weeks, and periodic
acid schiff's (PAS) reaction was used for detecting the expression of the liver glycogen after cultured in vitro for 4 weeks.Results:The results
showed that in 3D scaffold material, embryonic hepatocytes cultured in vitro could form some structures containing vascular-like
and liver -like tissues. AFP was highly expressed in liver-like tissues cultured for 1 weeks. ALB and liver glycogen were expressed in 3D
liver -like tissues cultured in vitro for 4 weeks.Conclusion:In 3D controlled assembly system, the embryonic hepatocytes which grow
three-dimensionally are able to formliver-like structures and show some liver functions. |
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