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Stem Cell Res Ther. 2015 Dec 12;6:246. doi: 10.1186/s13287-015-0227-6.

Transplantation of hESC-derived hepatocytes protects mice from liver injury.

Tolosa L1,2,3,4, Caron J5,6,7, Hannoun Z8,9,10, Antoni M11,12,13, López S14, Burks D15, Castell JV16,17, Weber A18,19,20, Gomez-Lechon MJ21,22, Dubart-Kupperschmitt A23,24,25.

Author information

1
INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. laiatolosa@hotmail.com.
2
Unidad de Hepatología Experimental, IIS LA Fe, Valencia, S-46026, Spain. laiatolosa@hotmail.com.
3
Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. laiatolosa@hotmail.com.
4
DHU Hepatinov, Villejuif, F-94800, France. laiatolosa@hotmail.com.
5
INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. jerome.caron@inserm.fr.
6
Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. jerome.caron@inserm.fr.
7
DHU Hepatinov, Villejuif, F-94800, France. jerome.caron@inserm.fr.
8
INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. zara.hannoun@inserm.fr.
9
Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. zara.hannoun@inserm.fr.
10
DHU Hepatinov, Villejuif, F-94800, France. zara.hannoun@inserm.fr.
11
INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. marc-antoni@hotmail.fr.
12
Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. marc-antoni@hotmail.fr.
13
DHU Hepatinov, Villejuif, F-94800, France. marc-antoni@hotmail.fr.
14
Unidad de Hepatología Experimental, IIS LA Fe, Valencia, S-46026, Spain. silvia_lopezcortes@hotmail.com.
15
CIBERDEM, Centro de Investigacion Prıncipe Felipe, Valencia, S-46012, Spain. dburks@cipf.es.
16
Unidad de Hepatología Experimental, IIS LA Fe, Valencia, S-46026, Spain. jose.castell@uv.es.
17
CIBERehd, FIS, Barcelona, S-08036, Spain. jose.castell@uv.es.
18
INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. anne.weber@inserm.fr.
19
Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. anne.weber@inserm.fr.
20
DHU Hepatinov, Villejuif, F-94800, France. anne.weber@inserm.fr.
21
Unidad de Hepatología Experimental, IIS LA Fe, Valencia, S-46026, Spain. gomez_mjo@gva.es.
22
CIBERehd, FIS, Barcelona, S-08036, Spain. gomez_mjo@gva.es.
23
INSERM, U 1193, Hôpital Paul Brousse, Villejuif, F-94807, France. anne.dubart@inserm.fr.
24
Univ Paris-Sud, UMR-S 1193, Villejuif, F-94800, France. anne.dubart@inserm.fr.
25
DHU Hepatinov, Villejuif, F-94800, France. anne.dubart@inserm.fr.

Abstract

BACKGROUND:

Hepatic cell therapy has become a viable alternative to liver transplantation for life-threatening liver diseases. However, the supply of human hepatocytes is limited due to the shortage of suitable donor organs required to isolate high-quality cells. Human pluripotent stem cells reflect a potential renewable source for generating functional hepatocytes. However, most differentiation protocols use undefined matrices or factors of animal origin; as such, the resulting hepatocytes are not Good Manufacturing Practice compliant. Moreover, the preclinical studies employed to assess safety and function of human embryonic stem cell (hESC)-derived hepatocytes are generally limited to immunodeficient mice. In the present study, we evaluate the generation of hepatocytes under defined conditions using a European hESC line (VAL9) which was derived under animal-free conditions. The function capacity of VAL9-derived hepatocytes was assessed by transplantation into mice with acetaminophen-induced acute liver failure, a clinically relevant model.

METHODS:

We developed a protocol that successfully differentiates hESCs into bipotent hepatic progenitors under defined conditions, without the use of chromatin modifiers such as dimethyl sulphoxide. These progenitors can be cryopreserved and are able to generate both committed precursors of cholangiocytes and neonate-like hepatocytes.

RESULTS:

Thirty days post-differentiation, hESCs expressed hepatocyte-specific markers such as asialoglycoprotein receptor and hepatic nuclear factors including HNF4α. The cells exhibited properties of mature hepatocytes such as urea secretion and UGT1A1 and cytochrome P450 activities. When transplanted into mice with acetaminophen-induced acute liver failure, a model of liver damage, the VAL9-derived hepatocytes efficiently engrafted and proliferated, repopulating up to 10 % of the liver. In these transplanted livers, we observed a significant decrease of liver transaminases and found no evidence of tumourigenicity. Thus, VAL9-derived hepatocytes were able to rescue hepatic function in acetaminophen-treated animals.

CONCLUSIONS:

Our study reveals an efficient protocol for differentiating VAL9 hESCs to neonatal hepatocytes which are then able to repopulate livers in vivo without tumour induction. The human hepatocytes are able to rescue liver function in mice with acetaminophen-induced acute toxicity. These results provide proof-of-concept that replacement therapies using hESC-derived hepatocytes are effective for treating liver diseases.

PMID:
26652177
PMCID:
PMC4676869
DOI:
10.1186/s13287-015-0227-6
[Indexed for MEDLINE]
Free PMC Article

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