Cell biology is different in small volumes: endogenous signals shape phenotype of primary hepatocytes cultured in microfluidic channels.
Cell biology is different in small volumes: endogenous signals shape phenotype of primary hepatocytes cultured in microfluidic channels.
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DOI：
10.1038/srep33980
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发表时间：
2016-09-29
影响因子：
4.6
通讯作者：
Revzin A
中科院分区：
文献类型：
--
作者：
Haque A;Gheibi P;Gao Y;Foster E;Son KJ;You J;Stybayeva G;Patel D;Revzin A
体外维持肝细胞的方法旨在通过使用共培养物、表面涂层和 3D 球体来概括天然肝脏微环境的各个方面。这项研究强调了空间限制的影响——体内微环境中研究较少的一个组成部分。我们证明，在低容量微流体通道（微室）中培养的肝细胞可保留分化的肝表型 21 天，而在相同条件下在常规培养板中培养的细胞在 7 天后去分化。仔细考虑营养输送和氧张力表明这些因素不能单独解释微室中细胞功能的增强。通过一系列涉及不同高度的微流体室和关键分子途径抑制的实验，我们证实小体积肝细胞的表型是由内源信号塑造的，包括肝细胞生长因子（HGF）等肝诱导生长因子（GF）和转化生长因子（TGF）-β1等肝破坏性GF。通常不认为肝细胞是 GF 的重要产生者——这种作用通常被分配给肝脏的非实质细胞。我们的研究表明，在适当的微环境中，肝细胞产生足以塑造其表型和功能的水平的肝诱导和促纤维化信号。
The approaches for maintaining hepatocytes in vitro are aimed at recapitulating aspects of the native liver microenvironment through the use of co-cultures, surface coatings and 3D spheroids. This study highlights the effects of spatial confinement-a less studied component of the in vivo microenvironment. We demonstrate that hepatocytes cultured in low-volume microfluidic channels (microchambers) retain differentiated hepatic phenotype for 21 days whereas cells cultured in regular culture plates under identical conditions de-differentiate after 7 days. Careful consideration of nutrient delivery and oxygen tension suggested that these factors could not solely account for enhanced cell function in microchambers. Through a series of experiments involving microfluidic chambers of various heights and inhibition of key molecular pathways, we confirmed that phenotype of hepatocytes in small volumes was shaped by endogenous signals, both hepato-inductive growth factors (GFs) such as hepatocyte growth factor (HGF) and hepato-disruptive GFs such as transforming growth factor (TGF)-β1. Hepatocytes are not generally thought of as significant producers of GFs–this role is typically assigned to nonparenchymal cells of the liver. Our study demonstrates that, in an appropriate microenvironment, hepatocytes produce hepato-inductive and pro-fibrogenic signals at the levels sufficient to shape their phenotype and function.
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DOI：
10.1111/j.1749-6632.1980.tb29518.x
发表时间：
1980-01-01
影响因子：
5.2
作者：
Bissell, D M;Guzelian, P S
通讯作者：
Guzelian, P S
影响因子：
6
作者：
Foster, Elena;You, Jungmok;Revzin, Alexander
通讯作者：
Revzin, Alexander
影响因子：
13.5
作者：
Inoue, Y;Tomiya, T;Fujiwara, K
通讯作者：
Fujiwara, K
影响因子：
2.9
作者：
DUNN, JCY;TOMPKINS, RG;YARMUSH, ML
通讯作者：
YARMUSH, ML
影响因子：
14
作者：
Haque, Amranul;Hexig, Bayar;Akaike, Toshihiro
通讯作者：
Akaike, Toshihiro