Efficient large-scale generation of functional hepatocytes from mouse embryonic stem cells grown in a rotating bioreactor with exogenous growth factors and hormones.
Efficient large-scale generation of functional hepatocytes from mouse embryonic stem cells grown in a rotating bioreactor with exogenous growth factors and hormones.
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DOI：
10.1186/scrt356
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发表时间：
2013
影响因子：
7.5
通讯作者：
Geng Y
中科院分区：
文献类型：
--
作者：
Zhang S;Zhang Y;Chen L;Liu T;Li Y;Wang Y;Geng Y
胚胎干细胞被认为是肝细胞移植和生物人工肝开发的潜在有利来源。然而，从ES细胞有效地大规模产生功能性肝细胞仍然是一个主要的挑战，特别是对于那些与临床应用兼容的方法。在这项研究中，我们研究了大量的功能性肝细胞是否可以从小鼠ES（mES）细胞分化使用模拟微重力生物反应器。在旋转生物反应器中，在外源生长因子和激素的存在下培养mES细胞以形成胚状体（EBs），然后胚状体分化为肝细胞。在旋转培养过程中，大部分EB源性细胞逐渐显示出正常肝细胞的组织学特征。更具体地，在来自生物反应器培养物的分化细胞中检测到比来自静态培养物的细胞中更高水平的肝基因和蛋白质的表达。在进一步生长，EB在组织培养板上，大多数EB衍生的细胞被发现显示肝细胞的形态特征，以及白蛋白的合成。此外，EB衍生细胞生长在旋转生物反应器中表现出更高水平的肝脏特异性功能，如糖原储存，细胞色素P450活性，低密度脂蛋白，和吲哚菁绿色吸收，比分化细胞生长在静态培养。当将来自第14天EB的EB衍生细胞和细胞培养上清液注射到裸鼠中时，移植的细胞被植入受体肝脏中。在旋转生物反应器中，mES细胞通过EB形成可以产生大量高质量的肝细胞。该系统可用于细胞移植和生物人工肝的开发中的肝细胞的大规模生成。
Embryonic stem (ES) cells are considered a potentially advantageous source of hepatocytes for both transplantation and the development of bioartificial livers. However, the efficient large-scale generation of functional hepatocytes from ES cells remains a major challenge, especially for those methods compatible with clinical applications. In this study, we investigated whether a large number of functional hepatocytes can be differentiated from mouse ES (mES) cells using a simulated microgravity bioreactor. mES cells were cultured in a rotating bioreactor in the presence of exogenous growth factors and hormones to form embryoid bodies (EBs), which then differentiated into hepatocytes. During the rotating culture, most of the EB-derived cells gradually showed the histologic characteristics of normal hepatocytes. More specifically, the expression of hepatic genes and proteins was detected at a higher level in the differentiated cells from the bioreactor culture than in cells from a static culture. On further growing, the EBs on tissue-culture plates, most of the EB-derived cells were found to display the morphologic features of hepatocytes, as well as albumin synthesis. In addition, the EB-derived cells grown in the rotating bioreactor exhibited higher levels of liver-specific functions, such as glycogen storage, cytochrome P450 activity, low-density lipoprotein, and indocyanine green uptake, than did differentiated cells grown in static culture. When the EB-derived cells from day-14 EBs and the cells’ culture supernatant were injected into nude mice, the transplanted cells were engrafted into the recipient livers. Large quantities of high-quality hepatocytes can be generated from mES cells in a rotating bioreactor via EB formation. This system may be useful in the large-scale generation of hepatocytes for both cell transplantation and the development of bioartificial livers.
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影响因子：
4
作者：
Schwartz, RE;Linehan, JL;Kaufman, DS
通讯作者：
Kaufman, DS
影响因子：
3.8
作者：
Gerecht-Nir, S;Cohen, S;Itskovitz-Eldor, J
通讯作者：
Itskovitz-Eldor, J
影响因子：
4.8
作者：
Cho, Cheul H.;Parashurama, Natesh;Yarmush, Martin L.
通讯作者：
Yarmush, Martin L.
影响因子：
4.1
作者：
Fridley, Krista M.;Fernandez, Irina;Roy, Krishnendu
通讯作者：
Roy, Krishnendu
影响因子：
3.3
作者：
Mizumoto, Hiroshi;Hayashi, Shunsuke;Kajiwara, Toshihisa
通讯作者：
Kajiwara, Toshihisa