Small Cell Colonies Appear in the Primary Culture of Adult Rat Hepatocytes in the Presence of Nicotinamide and Epidermal Growth Factor

Mitaka, Toshihiro; Mikami, Michihide; Sattler, Gerald L.; Pitot, Henry C.; Mochizuki, Yohichi

doi:10.1002/hep.1840160224

Cited by 115 publications

(92 citation statements)

References 31 publications

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“…Fetal hepatic cells were isolated from the porcine livers of embryos at 5 and 8 wk of gestation (NIBS strain) by the two-step liver perfusion method of Seglen with some modifications [15][16][17] . Five weeks of gestation was designated as E35 and 8 wk of gestation was designated as E56.…”

Section: Isolation Of Fetal Hepatocytes and Nonparenchymal Epithelialmentioning

confidence: 99%

Hepatic reconstruction from fetal porcine liver cells using a radial flow bioreactor

Ishii¹,

Saito²,

Marushima³

et al. 2008

WJG

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Section: Isolation Of Fetal Hepatocytes and Nonparenchymal Epithelialmentioning

confidence: 99%

Hepatic reconstruction from fetal porcine liver cells using a radial flow bioreactor

Ishii¹,

Saito²,

Marushima³

et al. 2008

WJG

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“…The medium was changed to fresh complete medium every 48 hrs. In this culture condition, hepatocytes maintain a fairly higher level of albumin and tryptophan 2,3-dioxygenase (TO) expression [11], but they enter the cell cycle and proliferate, and then colonies of small hepatocytes appear as described previously [12]. To further maintain mature hepatocyte phenotype, the next set of hepatocyte culture was conducted in collagen-1 gel sandwich configuration [13].…”

Section: Isolation and Culture Of Rat Hepatocytesmentioning

confidence: 99%

In Vitro Transdifferentiation of Mature Hepatocytes into Insulin-Producing Cells

et al. 2006

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Abstract. Adenovirus-mediated gene transfer of pancreatic duodenal homeobox transcription factor PDX-1, especially its super-active version (PDX-1/VP16), induces the expression of pancreatic hormones in murine liver and reverses streptozotocin-induced hyperglycemia. Histological analyses suggest that hepatocytes are the major source of insulinproducing cells by PDX-1 gene transfer, although the conversion of cultured hepatocytes into insulin-producing cells remains to be elucidated. The present study was conducted to address this issue. Hepatocytes were isolated from adult rats. Then, PDX-1 or PDX-1/VP16 gene was introduced by using adenovirus vector. Two days later, the expression of insulin was detected at mRNA and protein levels. Transfection of PDX-1/VP16 was more efficient in converting hepatocytes to insulin-producing cells. Immunoreactivity of albumin was downregulated in transdifferentiated cells and some of them almost completely lost albumin expression. During the course of transdifferentiation, upregulation of mRNA for CK19 and α-fetoprotein was observed. When cultured in collagen-1 gel sandwich configuration, hepatocytes maintained their mature phenotype and did not proliferate. In this condition, transfer of PDX-1/VP16 also induced the expression of insulin. These results clearly indicate that hepatocytes possess a potential to transdifferentiate into insulinproducing cells in vitro.

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“…[4][5][6][7][8] Other laboratories also reported that supplementation of the medium with nicotinamide and EGF stimulates the growth of the primary rat hepatocytes. 9,10 Most hepatocytes cultured in nicotinamide-supplemented medium can divide, and small hepatocytes (SHs), which are less than half as large as but are morphologically similar to MHs, proliferate so fast that the cells form colonies after 4 to 5 days of culture.…”

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confidence: 97%

Reconstruction of Hepatic Organoid by Rat Small Hepatocytes and Hepatic Nonparenchymal Cells

et al. 1999

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Landry et al. 1 first showed the reconstruction of a threedimensional cyto-architecture consisting of differentiated hepatocytes, bile duct-like cells, and deposited extracellular matrix (ECM) by the use of spheroidal aggregate culture of hepatic cells isolated from newborn rats. Thereafter, some attempts have been made to grow a hepatic organoid by the coculture of hepatocytes and fibroblasts. 2,3 As neither fibroblasts derived from the liver nor hepatocytes could proliferate, the size of the cell aggregates was limited and the position of the cells was disorderly, although differentiated functions were well maintained for a long time. To develop a hepatic organoid, the interactions not only between hepatocytes and nonparenchymal cells (NPCs) but also between ECM and hepatocytes must be well coordinated as many differentiated functions of mature hepatocytes (MHs), through which the homeostasis of life is sustained, should be maintained.We have reported that the proliferation of adult rat hepatocytes is observed in serum-free medium supplemented with 10 mmol/L nicotinamide and epidermal growth factor (EGF). [4][5][6][7][8] Other laboratories also reported that supplementation of the medium with nicotinamide and EGF stimulates the growth of the primary rat hepatocytes. 9,10 Most hepatocytes cultured in nicotinamide-supplemented medium can divide, and small hepatocytes (SHs), which are less than half as large as but are morphologically similar to MHs, proliferate so fast that the cells form colonies after 4 to 5 days of culture. They have been shown immunocytochemically and ultrastructurally to possess hepatic characteristics. 6 Recently, it was shown by our 11 and other laboratories 12 that a single SH can clonally proliferate and form a large colony consisting of more than 30 cells at day 10. The cells can grow and survive for more than 5 months. Thus, we previously proposed that rat hepatocytes may be classified into three types of cells with respect to their ability to divide: (1) cells that have a high potential to proliferate and form colonies in primary culture (type I cells; SHs), (2) cells for which the number of possible cell divisions is limited (type II cells), and (3) cells that lose the ability to divide (type III cells). 7,8 In addition, type II and type III cells may possess fully differentiated functions. Therefore, in this report we call both type II and type III cells MHs. Furthermore, we call the cell islands, which consist of SHs themselves and of a mixture of SHs and MHs, colonies.In the present experiment we showed that SHs could differentiate to MHs that interacted with hepatic NPCs and

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Small Cell Colonies Appear in the Primary Culture of Adult Rat Hepatocytes in the Presence of Nicotinamide and Epidermal Growth Factor

Cited by 115 publications

References 31 publications

Hepatic reconstruction from fetal porcine liver cells using a radial flow bioreactor

Hepatic reconstruction from fetal porcine liver cells using a radial flow bioreactor

In Vitro Transdifferentiation of Mature Hepatocytes into Insulin-Producing Cells

Reconstruction of Hepatic Organoid by Rat Small Hepatocytes and Hepatic Nonparenchymal Cells

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