Growth and albumin secretion of mouse fetal liver cells cryopreserved within porous polymer scaffolds as a viable cell source for bioartificial livers

Although cells isolated from fetal liver are one of the major sources for liver tissue engineering, it is still very difficult to induce them to fully differentiate in vitro into mature hepatocytes. We therefore investigated the effects of nicotinamide (NA), dimethyl sulfoxide (DMSO), and oncostatin M (OSM) on differentiation in terms of the expression of various liver-specific functions, because these factors have been reported to induce the emergence of possible hepatocyte progenitor cells (small hepatocytes) in adult rat hepatocyte culture or maturation of fetal mouse liver cells in culture. Fetal liver cells isolated from mouse embryos were cultured for 5 weeks in collagen-precoated plates. NA (10 mM) and DMSO (1%) remarkably enhanced the emergence of small hepatocytes, and OSM also synergistically enhanced the selective growth of small hepatocytes and inhibited the growth of blood cell populations. In the presence of these three factors, such small hepatocytes became dominant in culture, so that they covered almost 60–70% of confluence after week 2. In addition, some of them piled up over the small hepatocyte monolayer and displayed distinctively differentiated morphology, such as the emergence of binucleated cells, formation of tight gap junctions, and possible bile duct structures. Although OSM alone had very weak effects on hepatocyte functions, albumin secretion and cytochrome P450IA1/2 capacity were greatly enhanced when combined with NA or DMSO. This functional observation closely agreed with the emergence of small hepatocytes. In contrast, ammonium removal was strongly dependent on DMSO alone. DNA amount basis functions of fetal cells with three factors at week 5 were 1/7 for albumin secretion, 3 times higher for ammonium removal, and 1/10 for P450 capacity, compared with those of cultured adult mouse hepatocytes. These results show that inclusion of NA, DMSO, and OSM in the culture medium significantly enhances in vitro maturation of fetal liver cells when compared with conventional culture conditions.

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LONG-TERM CULTURE OF FETAL LIVER CELLS USING THREE-DIMENSIONAL POROUS POLYMER

ASAIO Journal ◽

10.1097/00002480-199903000-00045 ◽

1999 ◽

Vol 45 (2) ◽

pp. 129

Author(s):

H Miyoshi ◽

T Ehashi ◽

H Ema ◽

H C Hsu ◽

H Nakauchi ◽

...

Keyword(s):

Fetal Liver ◽

Three Dimensional ◽

Liver Cells ◽

Porous Polymer ◽

Long Term Culture ◽

Fetal Liver Cells

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Long-Term Culture of Fetal Liver Cells Using a Three-Dimensional Porous Polymer Substrate

ASAIO Journal ◽

10.1097/00002480-200007000-00005 ◽

2000 ◽

Vol 46 (4) ◽

pp. 397-402 ◽

Cited By ~ 26

Author(s):

Hirotoshi Miyoshi ◽

Tomo Ehashi ◽

Hideo Ema ◽

Hsiang Chun Hsu ◽

Hiromitsu Nakauchi ◽

...

Keyword(s):

Fetal Liver ◽

Three Dimensional ◽

Liver Cells ◽

Porous Polymer ◽

Polymer Substrate ◽

Long Term Culture ◽

Fetal Liver Cells

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Cultivation of Fetal Liver Cells in a Three-Dimensional Poly-L-Lactic Acid Scaffold in the Presence of Oncostatin M

Cell Transplantation ◽

10.3727/000000002783985648 ◽

2002 ◽

Vol 11 (5) ◽

pp. 403-406 ◽

Cited By ~ 27

Author(s):

Jinlan Jiang ◽

Nobuhiko Kojima ◽

Taisei Kinoshita ◽

Atsushi Miyajima ◽

Weiqun Yan ◽

...

Keyword(s):

Lactic Acid ◽

Fetal Liver ◽

Three Dimensional ◽

Liver Cells ◽

Oncostatin M ◽

Albumin Secretion ◽

Plla Scaffold ◽

Fetal Liver Cells ◽

Number Of Cells

To investigate the feasibility of fetal liver cells for liver tissue engineering, the supporting function of poly-l-lactic acid (PLLA) for fetal liver cells and the effects of oncostatin M (OSM) on hepatic differentiation were studied. After preparing three-dimensional biodegradable PLLA scaffold having a well-developed open-pore structure by a gas-forming method with ammonium chloride particles as a porogen and a gas-forming reagent, fetal liver cells separated from E14.5-C57BL/6CrSlc murine embryos were inoculated in the PLLA scaffolds. Cells were cultured in Williams' E medium with or without OSM (10 ng/ml) for 30 days with a medium change every 2 days. Results showed that there were significant increases in the number of cells and in albumin secretion in PLLA culture compared with in monolayer culture on day 15. In addition, a significant increase in albumin secretion was observed in OSM-added PLLA culture compared with OSM-free culture, and there was only a slightly enhanced albumin secretion in monolayer cultures with OSM. These results suggest that PLLA may enhance the biological activity of OSM for inducing maturation of fetal liver cells. Interestingly, the number of cells in PLLA culture with OSM decreased compared with OSM-free PLLA culture at day 15. This may be because promotion of hepatic development by OSM simultaneously suppressed in vitro hematopoiesis (i.e., blood cell production). In summary, our results indicate that the three-dimensional PLLA scaffold is a good support material for the cultivation of fetal liver cells and that OSM is capable of not only terminating hematopoiesis of the fetal liver but also stimulating the maturation of hepatic parenchymal cells in vitro.

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Influence of maternal diabetes in rats on hemoglobin synthesis and uridine uptake by fetal liver cells

Diabetes ◽

10.2337/diabetes.34.3.212 ◽

1985 ◽

Vol 34 (3) ◽

pp. 212-216

Author(s):

S. Mulay ◽

L. F. Congote

Keyword(s):

Fetal Liver ◽

Maternal Diabetes ◽

Liver Cells ◽

Hemoglobin Synthesis ◽

Uridine Uptake ◽

Fetal Liver Cells

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Use of human fetal liver cells for treatment of patients with lower limb peripheral artery disease

Cell and Organ Transplantology ◽

10.22494/cot.v2i1.39 ◽

2014 ◽

Vol 2 (1) ◽

pp. 10-13

Author(s):

R. Salyutin ◽

D. Dombrowski ◽

M. Komarov ◽

N. Sokolov ◽

S. Palyanitsya ◽

...

Keyword(s):

Progenitor Cells ◽

Lower Limb ◽

Fetal Liver ◽

Life Quality ◽

Liver Cells ◽

Surgical Reconstruction ◽

Clinical Effects ◽

Doppler Flowmetry ◽

Human Fetal Liver ◽

Fetal Liver Cells

In the group of patients (n = 21, mean age 54 ± 5.8 years) with chronic lower limb ischemia stage IIB who were non-liable for reconstructiverestoration surgery, we have established positive clinical effects of local transplantation of human fetal liver progenitor cells. Complex examination following 1, 3, 6 and 12 months after transplantation included duplex scanning of limb arteries, x-ray contrast arteriography and laser Doppler flowmetry as well as measuring pain-free walking and evaluating life quality based on individual questionnaire data.Owing to the transplant “Cryopreserved human fetal liver progenitor cells” the patients demonstrated stable increase of life quality index and pain-free walking as well as improvement of general health allowing assign them to the group of patients with lower ischemia stage, quicker social rehabilitation and lesser risk of disabling surgery (р < 0.05). Also, there were observations of improved microcirculation in the ischemic extremities owing to activation of endothelium-independent mechanisms of vasodilatation, reduced myotonus and neurotonus of the pre-capillaries and improved endothelium-dependent influence on the microhaemodynamic and, hence, an increased reserve capillary blood flow (p < 0.05).Analysis of the obtained results indicates prospects and effectiveness of using fetal liver cells transplantation in the patients who are not liable for surgical reconstruction of the vascular bed.

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Lower Expression of HNF4α and PGC1α Might Impair Rifampicin-mediated CYP3A4 Induction under Conditions Where PXR Is Overexpressed in Human Fetal Liver Cells

Drug Metabolism and Pharmacokinetics ◽

10.2133/dmpk.dmpk-11-rg-126 ◽

2012 ◽

Vol 27 (4) ◽

pp. 430-438 ◽

Cited By ~ 11

Author(s):

Takashi Takezawa ◽

Tamihide Matsunaga ◽

Kaori Aikawa ◽

Katsunori Nakamura ◽

Shigeru Ohmori

Keyword(s):

Fetal Liver ◽

Liver Cells ◽

Human Fetal Liver ◽

Fetal Liver Cells ◽

Human Fetal Liver Cells ◽

Lower Expression ◽

Cyp3a4 Induction

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Basis of hematopoietic defects in platelet-derived growth factor (PDGF)-B and PDGF β-receptor null mice

Blood ◽

10.1182/blood.v97.7.1990 ◽

2001 ◽

Vol 97 (7) ◽

pp. 1990-1998 ◽

Cited By ~ 48

Author(s):

Wolfgang E. Kaminski ◽

Per Lindahl ◽

Nancy L. Lin ◽

Virginia C. Broudy ◽

Jeffrey R. Crosby ◽

...

Keyword(s):

Growth Factor ◽

Fetal Liver ◽

Liver Cells ◽

Platelet Derived Growth Factor ◽

Wild Type ◽

Liver Growth ◽

Hematologic Disorders ◽

Fetal Liver Cells ◽

Β Receptor

Abstract Platelet-derived growth factor (PDGF)-B and PDGF β-receptor (PDGFRβ) deficiency in mice is embryonic lethal and results in cardiovascular, renal, placental, and hematologic disorders. The hematologic disorders are described, and a correlation with hepatic hypocellularity is demonstrated. To explore possible causes, the colony-forming activity of fetal liver cells in vitro was assessed, and hematopoietic chimeras were demonstrated by the transplantation of mutant fetal liver cells into lethally irradiated recipients. It was found that mutant colony formation is equivalent to that of wild-type controls. Hematopoietic chimeras reconstituted with PDGF-B−/−, PDGFRβ−/−, or wild-type fetal liver cells show complete engraftment (greater than 98%) with donor granulocytes, monocytes, B cells, and T cells and display none of the cardiovascular or hematologic abnormalities seen in mutants. In mouse embryos, PDGF-B is expressed by vascular endothelial cells and megakaryocytes. After birth, expression is seen in macrophages and neurons. This study demonstrates that hematopoietic PDGF-B or PDGFRβ expression is not required for hematopoiesis or integrity of the cardiovascular system. It is argued that metabolic stress arising from mutant defects in the placenta, heart, or blood vessels may lead to impaired liver growth and decreased production of blood cells. The chimera models in this study will serve as valuable tools to test the role of PDGF in inflammatory and immune responses.

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