Isolation, Culture, and Transplantation of Intrahepatic Biliary Epithelial Cells and Oval Cells

Sirica, Alphonse E.; Mathis, Georg A.; Sano, Nobuya; Elmore, Lynne W.

doi:10.1159/000163564

Cited by 121 publications

(74 citation statements)

References 50 publications

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“…Agents that selectively stimulate bile duct proliferation do not induce oval-cell proliferation. Induction of bile duct hyperplasia by nonhepatocarcinogens such as bile duct ligation, 4,4'-diaminodiphenylmethane (DDPM), or a-naphthylisothiocyanate (ANIT) do not induce AFP or albumin-containing duct cells (75)(76)(77). There is one report identifying AFP-containing bile duct cells after ANIT (78), but this was not found in our studies (75,76 (50,75,76).…”

Section: Bile Duct Proliferationmentioning

confidence: 61%

Cellular origin of cancer: dedifferentiation or stem cell maturation arrest?

Sell

1993

Environ Health Perspect

189

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Given the fundamental principle that cancer must arise from a cell that has the potential to divide, two major nonexclusive hypotheses of the cellular origin of cancer are that malignancy arises a) from stem cells due to maturation arrest or b) from dedifferentiation of mature cells that retain the ability to proliferate. The role of stem cells in carcinogenesis is clearly demonstrated in teratocarcinomas. The malignant stem cells of teratocarcinomas are derived from normal multipotent stem cells and have the potential to differentiate into normal benign mature tissue. A widely studied model supporting dedifferentiation has been the putative origin of hepatocarcinomas from "premalignant" foci and nodules induced in the rat liver by chemicals. However, the dedifferentiation concept for hepatocarcinogenesis is challenged by more recent interpretations indicating that hepatocellular carcinoma arises from maturation arrest caused by abherrant differentiation of determined stem cells. Either hypothesis is supported by the cellular changes that occur in the rodent liver after different hepatocarcinogenic regimens. The formation of foci and nodules from altered hepatocytes supports dedifferentiation; the proliferation of small oval cells with the potential to differentiate into either biliary ducts or hepatocytes supports arrested maturation of determined stem cells. It is now postulated that foci and nodular change reflect adaptive changes to the toxic effects of carcinogens and not "preneoplastic" stages to cancer. The stem cell model predicts that genotoxic chemicals induce mutations in the determined stem cell which may be expressed in its progeny. Proliferation of initiated cells is induced by promoting events which also allow additional mutations to occur.

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Section: Bile Duct Proliferationmentioning

confidence: 61%

Cellular origin of cancer: dedifferentiation or stem cell maturation arrest?

Sell

1993

Environ Health Perspect

189

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“…Proliferation of small AFP ϩ oval cells was first observed in primary cultures of liver cells isolated by gravity sedimentation from rats fed AAFsupplemented choline-deficient diets 2 ; similar results were reported with other such models. [28][29][30] Starting with rats fed ethionine in choline-deficient diets, 31 oval cell cultures were obtained that produced transformed cells after prolonged culture, 32,33 after transfection with oncogenes, [33][34][35] and after mutagenesis. [36][37][38] Many of these lines produced tumors when injected into nude mice.…”

Section: Discussionmentioning

confidence: 99%

Derivation, characterization, and phenotypic variation of hepatic progenitor cell lines isolated from adult rats

Sun

Ilić

et al. 2002

Hepatology

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Liver progenitor cells (LPCs) cloned from adult rat livers following allyl alcohol injury express hematopoietic stem cell and early hepatic lineage markers when cultured on feeder layers; under these conditions, neither mature hepatocyte nor bile duct, Ito, stellate, Kupffer cell, or macrophage markers are detected. These phenotypes have remained stable without aneuploidy or morphological transformation after more than 100 population doublings. When cultured without feeder layers, the early lineage markers disappear, and mature hepatocyte markers are expressed; mature hepatocytic differentiation and cell size are also augmented by polypeptide and steroidal growth factors. In contrast to hepatocytic potential, duct-like structures and biliary epithelial markers are expressed on Matrigel. Because they were derived without carcinogens or mutagens, these bipotential LPC lines provide novel tools for models of cellular plasticity and hepatocarcinogenesis, as well as lines for use in cellular transplantation, gene therapy, and bioreactor construction. Tissue determined LPCs may reside in or near the terminal biliary tree, but it has been difficult to identify authentic LPCs in the normal liver. 2,3 One of the principle lines of evidence for existence of LPCs is appearance of oval cells after different kinds of liver injury, which differentiate into either hepatocytes or cholangiocytes. For example, using carcinogenic regimens where N-2-acetylaminofluorene (AAF) inhibits proliferation of mature hepatocytes, 4,5 oval cells appear in large numbers in duct-like structures. 6,7 Similarly, marked expansions of ductular oval cells occur when centrolobular injury is induced by CCl 4 and bile ductular hyperplasia is stimulated by bile duct ligation 8 or hepatocyte proliferation is inhibited by AAF. 9,10 Although these observations suggest that LPCs exist within biliary ducts, other models implicate nonductal cells in the portal triad. 11 Thus, nondescript periductular oval cells proliferate within 2 days after allyl alcohol (AA)-induced periportal injury, and 2 to 4 days later the periportal necrotic zone is replaced by an expanding population of oval cells and transitional hepatocytes, with little ductular reaction. 12-14 Based on these and earlier observations, 2 we postulated that nontransformed LPC lines, derived from oval cells induced after AA injury, might exhibit different stages of liver-cell differentiation and bipotential capacities of differentiation. We now describe the isolation and characterization of 11 LPC strains and lines from adult livers after periportal injury induced by AA. Materials and Methods Materials.The following materials were obtained commercially: dexamethasone and bFGF (Sigma); Oncostatin M (R&D Systems, Minneapolis, MN); Matrigel (Collaborative Research, Bedford, MA); human epidermoid carcinoma cell line A431 lysates (Santa Cruz Biotechnology, Santa Cruz, CA).Derivation, Culture, and Storage of LPC Strains. Nineweek-old Fisher/344 female rats weighing 140-150 g were obtained from Harlan, India...

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“…Lines of liver epithelial (oval) cells have been developed which may serve as hepatocyte progenitors but differ morphologically from mature hepatocytes (29,(34)(35)(36). Recently it has been shown that human liver epithelial cells immortalized with simian virus 40 T antigen can express xenobiotic drugmetabolizing enzymes (37).…”

mentioning

confidence: 99%

Establishment and characterization of differentiated, nontransformed hepatocyte cell lines derived from mice transgenic for transforming growth factor alpha.

Merlino

Fausto

1994

Proc. Natl. Acad. Sci. U.S.A.

267

199

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Hepatocytes are extensively used in studies of gene regulation but cannot be maintained in long-term culture as replicating, differentiated cells while remaining nontumorigenic. We have derived two hepatocyte lines from livers of transgenic mice overexpressing transforming growth factor a, a potent hepatocyte mitogen, which overcome these limitations. The transgenic hepatocytes were maintained for .2 months in serum-supplemented primary culture and gave rise to cell lines, of which two (AML12 and AML14) have been cultured for >1.5 years (>80 passages). Both lines have typical hepatocyte features such as peroxisomes and bile canalicular-like structures, do not grow in soft agar, and are nontumorigenic in nude mice. Like normal hepatocytes, AML cells express high levels of mRNA for serum (albumin, al-antitrypsin, and transferrin) and gap junction (connexins 26 and 32) proteins, secrete albumin, and contain solely isozyme 5 of lactate dehydrogenase. After extensive passaging, AML12 cells continue to strongly coexpress hepatocyte connexin mRNAs but do not display nonparenchymal cell markers. Although mRNA levels for some serum proteins progressively fall, high expression in late AML12 cultures may be regained by passage in serum-free medium. The AML14 line loses expression of both differentiated markers and transgene mRNA with extended passaging, and hepatocytic traits are only partially restored by passage in serum-free medium. These differentiated, nontumorigenic cell lines should serve as models in which to study hepatocyte growth and differentiation.Mammalian hepatocytes have long been used to investigate mechanisms of cell growth, differentiation, and tumorigenesis and as vectors for gene therapy. These studies have been hampered because dissociated hepatocytes rapidly lose their cell-specific functions, are short-lived, and have a limited ability to replicate in culture. Longer-term differentiated cultures can be maintained with specialized media and substrata or by coculture with nonparenchymal cells, but under these conditions the cells remain nonproliferative (1-3).Hepatocyte lines which express some tissue-specific markers have been propagated in vitro from hepatocellular tumors. More recently, well-differentiated lines have been derived by introducing viral oncogenes into hepatocytes. While useful, these lines have the drawback of either being fully transformed or undergoing transformation with repeated passaging (3)(4)(5)(6).Transforming growth factor a (TGFa) is a polypeptide that regulates normal growth in epithelial tissues, and its overproduction in cells possessing epidermal growth factor receptors (EGFRs) is often correlated with malignant transformation (7-10). TGFa is expressed in the liver during embryogenesisThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. and transiently increases during liver regeneration after partial hepa...

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Isolation, Culture, and Transplantation of Intrahepatic Biliary Epithelial Cells and Oval Cells

Cited by 121 publications

References 50 publications

Cellular origin of cancer: dedifferentiation or stem cell maturation arrest?

Cellular origin of cancer: dedifferentiation or stem cell maturation arrest?

Derivation, characterization, and phenotypic variation of hepatic progenitor cell lines isolated from adult rats

Establishment and characterization of differentiated, nontransformed hepatocyte cell lines derived from mice transgenic for transforming growth factor alpha.

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