Heparin-based hydrogel as a matrix for encapsulation and cultivation of primary hepatocytes.
Heparin-based hydrogel as a matrix for encapsulation and cultivation of primary hepatocytes.
复制标题
DOI：
10.1016/j.biomaterials.2010.01.068
复制
发表时间：
2010-05
期刊：
影响因子：
14
通讯作者：
Tae, Giyoong
中科院分区：
文献类型：
--
作者：
Kim, Mihye;Lee, Ji Youn;Jones, Caroline N.;Revzin, Alexander;Tae, Giyoong
原代肝细胞通常用作毒理学和组织工程领域的肝脏替代物，因此，体外维持功能性肝细胞是一个重要的研究课题。本文试图将基于肝素的水凝胶作为肝细胞培养的三维支架进行表征。将原代大鼠肝细胞与由硫醇化肝素和丙烯酸酯化聚乙二醇（PEG）组成的预聚物溶液混合。将温度从 25°C 升高至 37°C 引发硫醇和丙烯酸酯部分之间的迈克尔加成反应，并导致形成带有截留细胞的水凝胶。对肝脏特异性产物白蛋白和尿素的分析表明，肝素水凝胶对细胞无细胞毒性，实际上可以促进肝功能。培养三周后，包裹在肝素水凝胶中的肝细胞仍保持高水平的白蛋白和尿素合成。由于已知肝素可以结合生长因子，因此我们将肝细胞生长因子（HGF）——一种重要的肝脏信号分子——融入到水凝胶中。使用酶联免疫测定 (ELISA) 分析了肝素水凝胶基质中 HGF 的释放，结果显示，HGF 以受控方式发生，培养 30 天后仅释放 40% 的 GF 分子。重要的是，与单独在水凝胶中培养的细胞相比，在含有 HGF 的水凝胶中培养的肝细胞表现出显着更高水平的白蛋白和尿素合成。总体而言，基于肝素的水凝胶被证明是一种有前途的基质，用于封装和维持难以培养的原代肝细胞。未来，我们设想使用基于肝素的水凝胶作为肝细胞或干细胞体外分化的基质以及作为这些细胞移植的载体。
Primary hepatocytes are commonly used as liver surrogates in toxicology and tissue engineering fields, therefore, maintenance of functional hepatocytes in vitro is an important topic of investigation. This paper sought to characterize heparin-based hydrogel as a three-dimensional scaffold for hepatocyte culture. The primary rat hepatocytes were mixed with a prepolymer solution comprised of thiolated heparin and acrylated poly(ethylene glycol) (PEG). Raising the temperature from 25° to 37°C initiated Michael addition reaction between the thiol and acrylated moieties and resulted in formation of hydrogel with entrapped cells. Analysis of liver-specific products, albumin and urea, revealed that the heparin hydrogel was non-cytotoxic to cells and, in fact, promoted hepatic function. Hepatocytes entrapped in the heparin-based hydrogel maintained high levels of albumin and urea synthesis after three weeks in culture. Because heparin is known to bind growth factors, we incorporated hepatocyte growth factor (HGF) – an important liver signaling molecule - into the hydrogel. HGF release from heparin hydrogel matrix was analyzed using enzyme linked immunoassay (ELISA) and was shown to occur in a controlled manner with only 40% of GF molecules released after 30 days in culture. Importantly, hepatocytes cultured within HGF-containing hydrogels exhibited significantly higher levels of albumin and urea synthesis compared to cells cultured in the hydrogel alone. Overall, heparin-based hydrogel showed to be a promising matrix for encapsulation and maintenance of difficult-to-culture primary hepatocytes. In the future, we envision employing heparin-based hyrogels as matrices for in vitro differentiation of hepatocytes or stem cells and as vehicles for transplantation of these cells.
登录
查看更多内容
影响因子：
3.7
作者：
KATO, S;ISHII, T;AKAMATSU, N
通讯作者：
AKAMATSU, N
影响因子：
--
作者：
Bhandari, RNB;Riccalton, LA;Shakesheff, KM
通讯作者：
Shakesheff, KM
影响因子：
2.9
作者：
DUNN, JCY;TOMPKINS, RG;YARMUSH, ML
通讯作者：
YARMUSH, ML
影响因子：
6.2
作者：
Choi, Won Il;Kim, Mihye;Kim, Young Ha
通讯作者：
Kim, Young Ha
影响因子：
3.3
作者：
Fassett, J;Tobolt, D;Hansen, LK
通讯作者：
Hansen, LK