Naslov
In Vivo Pharmacological Selection Increases by 50-fold Transplanted Human Hepatocyte Survival in Immunocompetent Rats

Autori
Madadi, Shilpa ; Volarević, Martina ; Smolić, Robert ; Coash, Marcy ; Smith, Jeanette ; Andorfer, John ; Kelly, Penny ; Wu, Catherine ; Wu, George

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Hepatology / Keith D. Lindor - Hoboken (NJ) : John Wiley & Sons, 2008, 1123A-1123A

Skup
59th Annual Meeting of the American Association for the Study of Liver Diseases, San Francisco 2008

Mjesto i datum
San Francisco (CA), Sjedinjene Američke Države, 31.10.2008. - 04.11.2008

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
jetra ; hepatociti ; imunokompetentni štakori ; siRNA ; GFP-Huh 7 stanice ; CYP2E1 gen
(liver ; hepatocytes ; immunocompetant rats ; siRNAs ; GFP-Huh 7 cells ; CYP2E1 gene)

Sažetak
Purpose: We have shown that transplanted human hepatocytes can survive in immunocompetant rats when hosts are rendered tolerant by intrafetal inoculation of those cells [Gastroenterol. 2006 128:1416-1423]. However, the fraction of human cells in this model did not exceed 1%. To increase the ratio of human to rat liver cells, human hepatocytes with a survival advantage over host cells were transplanted and hepatotoxic doses of a selection agent administered in situ. Methods: siRNAs were prepared to down-regulate CYP2E1 which converts acetaminophen (APAP) into toxic metabolites. However, GFP-Huh 7 cells without siRNA transfection had low levels of CYP2E1 RNA, and this conferred protection against APAP in cell culture compared to rat hepatocytes. Fetal rats were tolerized with GFP-Huh7 cells, and 5 x 106 GFP-Huh 7 cells were transplanted 5 days after birth by splenic injection. Rats were treated with APAP 1250 mg/kg (n=13) or normal saline (n=2) by intraperitoneal injection on day 19. ALT was measured 1 day prior and on days 1-7 after injection. BrdU was injected 1 day prior to sacrifice. Livers from treated and control rats were stained with anti-BrdU, anti-human albumin, DAPI, and examined by histology, and immunohistochemistry visualized by fluorescence microscopy. Results: As expected, ALT peaked 1-2 days after APAP injection with means of 180 U/l, while saline-injected controls had means of 75 U/l. See 'Human hepatocyte number, GFP positive cells in APAP-treated rats' table. On day 1, 2-3 GFP (+) cells were noted in each field. However, on day 7, 40-50 GFP (+) cells in clusters were seen in each field. There were no GFP (+) cells in the control non-APAP-treated rats. Human albumin was present in GFP (+) cells, and the number of dividing cells in APAP-treated rats was greatly increased compared to controls on day 7. Conclusions: Low CYP2E1 gene expression in Huh-7 cells provides an effective survival advantage in the presence of APAP in vitro. That difference between Huh-7 and host hepatocytes provides an efficient means for pharmacological selection resulting in >50-fold enrichment in rats. Down regulation of CYPs by siRNA may permit application of the selection to other liver cell lines. Transplanted Huh-7 cells respond to regenerative stimuli from APAP-induced liver damage with substantial proliferation in rats.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

Napomena
Druge sekundarne publikacije koje indeksiraju časopis: * Biological Abstracts® (Thomson ISI) * BIOSIS Previews® (Thomson ISI) * CAB Abstracts® (CABI) * Chemical Abstracts Service/SciFinder (ACS) * Current Awareness in Biological Sciences (Elsevier) * Current Contents® /Clinical Medicine (Thomson ISI) * Current Contents® /Life Sciences (Thomson ISI) * EMBASE/Excerpta Medica (Elsevier) * ETOH (National Institute on Alcohol Abuse and Alcoholism) * Index Medicus/MEDLINE/PubMed (NLM) * International Pharmaceutical Abstracts (Thomson Scientific) * Journal Citation Reports/Science Edition (Thomson ISI) * Reference Update (Thomson ISI) * Science Citation Index Expanded™ (Thomson ISI) * Science Citation Index® (Thomson ISI) * SCOPUS (Elsevier) * SIIC Databases (Sociedad Iberoamericana de Informacion Cientifica) * Web of Science® (Thomson ISI)

POVEZANOST RADA