ESTABLISHMENT OF AN IMMORTALIZED HUMAN-LIVER ENDOTHELIAL CELL LINE WITH SV40T AND hTERT

Matsumura, Tatsuro; Takesue, Michihiko; Westerman, Karen A.; Okitsu, Teru; Sakaguchi, Masakiyo; Fukazawa, Takuya; Totsugawa, Toshinori; Noguchi, Hirofumi; Yamamoto, Shinichiro; Stolz, Donna B.; Tanaka, Noriaki; Leboulch, Philippe; Kobayashi, Naoya

doi:10.1097/01.tp.0000124286.82961.7e

Cited by 77 publications

(72 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We devised a strategy for differentiation of ES cells into hepatocyte-like cells using a four-step differentiation protocol: (i) generation of EBs; (ii) induction of definitive endoderm from 2-d-old EBs by growth in Activin A and FGF-2; (iii) induction of hepatic progenitors by co-culture with human liver nonparenchymal cell lines (human liver endothelial (TMNK-1) 14 , stellate (TWNT-1) 15 and cholangiocyte (MMNK-1) 16 cell lines) and a deleted variant of HGF (dHGF); and (iv) maturation into hepaticlike cells by culture in DMSO and dexamethasone (Fig. 1).…”

Section: Protocols Have Been Developed To Differentiate and Enrich Vamentioning

confidence: 99%

“…Induction of hepatic progenitor cells (expressing both albumin and AFP) and maturation into hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines 12| Prepare human liver nonparenchymal cell lines [14][15][16] , which can be maintained in T75 tissue culture flasks in 10 ml of DMEM and 10% FBS. Passage confluent cultures every week, and change the medium every 3 d.…”

Section: |mentioning

confidence: 99%

See 1 more Smart Citation

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

et al. 2007

Self Cite

View full text Add to dashboard Cite

This protocol describes a co-culture system for the in vitro differentiation of mouse embryonic stem cells into hepatocyte-like cells. Differentiation involves four steps: (i) formation of embryoid bodies (EB), (ii) induction of definitive endoderm from 2-d-old EBs, (iii) induction of hepatic progenitor cells and (iv) maturation into hepatocyte-like cells. Differentiation is completed by 16 d of culture. EBs are formed, and cells can be induced to differentiate into definitive endoderm by culture in Activin A and fibroblast growth factor 2 (FGF-2). Hepatic differentiation and maturation of cells is accomplished by withdrawal of Activin A and FGF-2 and by exposure to liver nonparenchymal cell-derived growth factors, a deleted variant of hepatocyte growth factor (dHGF) and dexamethasone. Approximately 70% of differentiated embryonic stem (ES) cells express albumin and can be recovered by albumin promoter-based cell sorting. The sorted cells produce albumin in culture and metabolize ammonia, lidocaine and diazepam at approximately two-thirds the rate of primary mouse hepatocytes.

show abstract

Section: Protocols Have Been Developed To Differentiate and Enrich Vamentioning

confidence: 99%

Section: |mentioning

confidence: 99%

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

et al. 2007

Self Cite

View full text Add to dashboard Cite

show abstract

“…1D). Coencapsulation of the human liver endothelial cell (LEC) line TMNK-1 (28,29) was the most beneficial to rat hepatocellular function ( Fig. 1 E and F), whereas additional mouse FIB were moderately and transiently supportive, and the human hepatic stellate cell (HSC) line TWNT-1 (30) had no effect (Fig.…”

Section: Resultsmentioning

confidence: 99%

Humanized mice with ectopic artificial liver tissues

Chen

Thomas

Ong

et al. 2011

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

144

148

View full text Add to dashboard Cite

"Humanized" mice offer a window into aspects of human physiology that are otherwise inaccessible. The best available methods for liver humanization rely on cell transplantation into immunodeficient mice with liver injury but these methods have not gained widespread use due to the duration and variability of hepatocyte repopulation. In light of the significant progress that has been achieved in clinical cell transplantation through tissue engineering, we sought to develop a humanized mouse model based on the facile and ectopic implantation of a tissue-engineered human liver. These human ectopic artificial livers (HEALs) stabilize the function of cryopreserved primary human hepatocytes through juxtacrine and paracrine signals in polymeric scaffolds. In contrast to current methods, HEALs can be efficiently established in immunocompetent mice with normal liver function. Mice transplanted with HEALs exhibit humanized liver functions persistent for weeks, including synthesis of human proteins, human drug metabolism, drug-drug interaction, and drug-induced liver injury. Here, mice with HEALs are used to predict the disproportionate metabolism and toxicity of "major" human metabolites using multiple routes of administration and monitoring. These advances may enable manufacturing of reproducible in vivo models for diverse drug development and research applications.P reclinical models provide a critical window into human physiology and are important for determining drug safety and efficacy. Bioengineering advances have produced in vivo platforms with improved disease modeling (1, 2), absorption, distribution, metabolism, excretion, and toxicity (ADME/Tox) (3, 4), and circulation-mimicking capabilities (4). "Human-on-a-chip" systems, however, show limited recapitulation of pharmacokinetics and cannot support clinically relevant administration routes. Elsewhere, humanized mice, based on genetic manipulation or transplantation of human cells, have also been highly enabling (5). Mice with chimeric livers show encouraging drug responsiveness (6, 7) and pathogen susceptibility (8-12). However, these models are confined to liver-injury, immunodeficient recipients and depend on the coordinated injection of high-quality human hepatocytes, which must home to the liver from the injection site, engraft, and expand over weeks to months within the injured host (13,14). Given the limitations of current preclinical models, human metabolites and their downstream effects often go undetected until clinical trials, the most costly and dangerous phase of drug development (15).Here we develop an improved humanized mouse model by implantation of tissue-engineered human ectopic artificial livers (HEALs). In the field of tissue engineering, polymeric scaffolds are widely used for cell and drug delivery in vivo (16,17). Primary hepatocytes, however, are challenging to maintain and implant via biomaterials due to their unstable phenotype, variable engraftment efficiencies, and high metabolic needs (18). Use of partial hepatectomy with portal-caval ...

show abstract

“…We used TMNK-1 cells (immortalized human LSECs), instead of primary LSECs, since their utility and efficiency was confirmed in the previous basic research (Matsumura et al, 2004).…”

Section: The Effect With Liver Sinusoidal Endothelial Cellsmentioning

confidence: 99%

Platelet and Liver Regeneration

Ohkohchi¹,

Murata²,

Takahashi³

2012

Tissue Regeneration - From Basic Biology to Clinical Application

View full text Add to dashboard Cite

ESTABLISHMENT OF AN IMMORTALIZED HUMAN-LIVER ENDOTHELIAL CELL LINE WITH SV40T AND hTERT

Abstract: This study demonstrates the utility and efficiency of the reversible immortalization procedure to expand primary human LECs for basic studies.

Cited by 77 publications

References 30 publications

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

Humanized mice with ectopic artificial liver tissues

Platelet and Liver Regeneration

Contact Info

Product

Resources

About