Effect of Collagen Gel Configuration on the Cytoskeleton in Cultured Rat Hepatocytes

Ezzell, Robert M.; Toner, Mehmet; Hendricks, Kristin B.; Dunn, James C.Y.; Tompkins, Ronald G.; Yarmush, Martin L.

doi:10.1006/excr.1993.1266

Cited by 96 publications

(74 citation statements)

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“…This is in accordance with reports on HCs that required no extensive matrix deposition when cultured in a 3-dimensional matrix scaffold. 50 Thus, results from our coculture experiments clearly indicate that primary, mature biliary epithelial cells from normal and cirrhotic liver depend not on HGF for bile duct generation. Instead, human BECs produce potent factor(s) other than HGF, which regulate differentiated morphogenesis of the biliary epithelium.…”

Section: Discussionmentioning

confidence: 58%

“…In our model, we find few if any nonparenchymal cells (possible sources for laminin deposition) other than BECs and HCs inside collagen DG secrete very little fibronectin or laminin. 50 Because cocultured HCs and BECs exhibited very little reticulin fibers in our system and ultrastructurally no basement membrane was detected around ductular structures FIG. 7.…”

Section: Discussionmentioning

confidence: 93%

“…Although it is clear that HGF is not the active principle, we have not yet identified the specific factor(s) present in CM that are responsible. Apart from the possible release of smaller amounts of soluble ECM components by BECs 45,49 or by HCs, 47,49,50 production of growth factors or cytokines by one or both of the epithelial cell types is implied. Potential growth factors produced by liver epithelial cells comprise IGF-1, which is mitogenic for stellate cells 42 and BECs, 32 FGF-1, and TGF-␣, which act in an autocrine fashion on HCs.…”

Section: Discussionmentioning

confidence: 99%

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Morphogenesis of primary human biliary epithelial cells: Induction in high-density culture or by coculture with autologous human hepatocytes

et al. 2001

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Although the control of biliary ductular morphogenesis has received some attention particularly using isolated rat biliary epithelial cell models, the regulation of human bile duct formation is not well defined. In the present study, using a 3-dimensional culture model comprising primary human biliary epithelial cells (BECs) and coculture with primary human hepatocytes, we have sought to define the factors involved. We have shown that primary human BECs can be expanded on collagen gels in the absence of growth factors or serum. When plated in high density in double collagen gels, BECs established 3-dimensional structures that subsequently developed into well differentiated polarized luminal ducts. This morphogenic response occurred in the absence of hepatocyte growth factor (HGF) and epidermal growth factor. Strikingly, the addition of growth factors While the control of liver regeneration has focused on the factors that regulate the initiation of growth in primary hepatocytes (HCs) 1,2 since they are the first cell type in the liver to respond in the regenerative process, 3 less attention has been paid to the biliary epithelial cell (BEC) compartment. We know that in vivo, BECs respond to partial hepatectomy with a delay of approximately 24 hours 4 (together with the sinusoidal cell populations). Although some of the mitogenic signaling molecules have been identified, 1,3 the factors that induce biliary ductular morphogenesis, essential to the efficient regeneration of the bile drainage system, are not well characterized.Many rodent-based biliary epithelial cell studies have focused on the bile duct ligation model, which results in extensive hyperplasia of ductular epithelium and indeed is a chosen means of increasing the yield of BECs that can be isolated from liver tissue. [5][6][7] In rats, factors including epidermal growth factor (EGF), hepatocyte growth factor (HGF), somatostatin, bile acids, and cytokines have been implicated in the proliferative response 6-10 after bile duct ligation 6,7 or partial hepatectomy. 10 After bile duct ligation in mice, HGF receptors (cmet) are up-regulated and interleukin 6 (IL-6) receptors (gp-80) are induced on the proliferating ductular cells. 11 Studies on isolated intrahepatic BECs from normal rats have also proven productive. 8,12,13 Other investigators have used dissected rodent bile duct units to show that interactions of the periductal inflammatory stroma are the source of these factors 11 and to perform functional studies on intrahepatic biliary epithelium. 12,[14][15][16][17][18] Some work on the control of ductular morphogenesis in vitro has also been performed with BECs from bile ductligated rats 5,6,18 and normal rats, 15-17 but the important regulatory factors remain poorly characterized. In other systems, most notably during angiogenesis and mammary gland ductulogenesis, factors such as EGF and HGF have been shown to be the principal morphogenic factors. [19][20][21][22][23][24] Recent evidence from in vitro-based cell proliferation and motility assays has ...

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Section: Discussionmentioning

confidence: 58%

Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

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Morphogenesis of primary human biliary epithelial cells: Induction in high-density culture or by coculture with autologous human hepatocytes

et al. 2001

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“…The collagen "gel sandwich" configuration is a popular method for maintaining long-term hepatocyte functions. [251][252][253][254][255][256][257][258][259] Nonetheless, the successful development of cell-based artificial liver devices requires long-term, stable hepatocytic function in a perfusable environment and efficient transport of nutrients and waste within the device.…”

Section: Vf Liver Cell Biology On a Chipmentioning

confidence: 99%

Biology on a Chip: Microfabrication for Studying the Behavior of Cultured Cells

Tourovskaia

Folch

2003

Crit Rev Biomed Eng

173

140

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The ability to culture cells in vitro has revolutionized hypothesis testing in basic cell and molecular biology research and has become a standard methodology in drug screening and toxicology assays. However, the traditional cell culture methodology-consisting essentially of the immersion of a large population of cells in a homogeneous fluid medium-has become increasingly limiting, both from a fundamental point of view (cells in vivo are surrounded by complex spatiotemporal microenvironments) and from a practical perspective (scaling up the number of fluid handling steps and cell manipulations for high-throughput studies in vitro is prohibitively expensive). Micro fabrication technologies have enabled researchers to design, with micrometer control, the biochemical composition and topology of the substrate, the medium composition, as well as the type of neighboring cells surrounding the microenvironment of the cell. In addition, microtechnology is conceptually well suited for the development of fast, low-cost in vitro systems that allow for high-throughput culturing and analysis of cells under large numbers of conditions. Here we review a variety of applications of microfabrication in cell culture studies, with an emphasis on the biology of various cell types.

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“…As growth on extracellular matrix (ECM) layers has been shown to enhance the expression of hepatocyte-specific differentiation markers [36][37][38][39][40][41], we investigated the effect of cultivating HepG2.2.15 cells on collagen I from rat tail. Figure 4 shows that HBV DNA expression in the supernatant increased from 5.0 !…”

Section: Effect Of Collagenmentioning

confidence: 99%

Optimised Conditions for the Production of Hepatitis B Virus from Cell Culture

Glebe¹,

Berting²,

Broehl³

et al. 2001

Intervirology

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In chronically infected patients, hepatitis B virus (HBV) particles reach numbers as large as >10⁹ genome equivalents (GE)/ml of serum. However, expression of infectious HBV particles in cell culture only yields 10⁵–10⁶ GE/ml, which is insufficient for many studies. HBV transcription and possibly replication is dependent on hepatocyte-specific differentiation. Thus, we tested several cell culture parameters that have been reported to enhance the expression of hepatocyte-specific markers, such as growth on different extracellular matrices, different cell culture media, low concentrations of fetal calf serum (FCS) and the addition of dimethyl sulfoxide (DMSO) to the medium. Lower concentrations of FCS, growth on collagen and inclusion of DMSO in the medium only moderately enhanced HBV production in vitro when applied individually. However, combinations of these parameters optimised cell culture conditions and reproducibly increased the release of HBV particles about 100-fold to titres >10⁸ GE/ml of culture medium.

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Effect of Collagen Gel Configuration on the Cytoskeleton in Cultured Rat Hepatocytes

Cited by 96 publications

References 0 publications

Morphogenesis of primary human biliary epithelial cells: Induction in high-density culture or by coculture with autologous human hepatocytes

Morphogenesis of primary human biliary epithelial cells: Induction in high-density culture or by coculture with autologous human hepatocytes

Biology on a Chip: Microfabrication for Studying the Behavior of Cultured Cells

Optimised Conditions for the Production of Hepatitis B Virus from Cell Culture

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