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Stem Cell Res Ther. 2015 Dec 1;6:237. doi: 10.1186/s13287-015-0218-7.

Hypoxic preconditioning potentiates the trophic effects of mesenchymal stem cells on co-cultured human primary hepatocytes.

Author information

1
Dhawan Lab, Institute of Liver Studies, King's College London and King's College Hospital NHS Foundation Trust, London, SE5 9PJ, UK. hong.qin@outlook.com.
2
Dhawan Lab, Institute of Liver Studies, King's College London and King's College Hospital NHS Foundation Trust, London, SE5 9PJ, UK. celine.filippi@kcl.ac.uk.
3
NIHR Biomedical Research Centre at Guy's and St Thomas NHS Foundation Trust and King's College London, London, SE1 9RT, UK. celine.filippi@kcl.ac.uk.
4
Dhawan Lab, Institute of Liver Studies, King's College London and King's College Hospital NHS Foundation Trust, London, SE5 9PJ, UK. sunsongmed@163.com.
5
Dhawan Lab, Institute of Liver Studies, King's College London and King's College Hospital NHS Foundation Trust, London, SE5 9PJ, UK. sharon.lehec@nhs.net.
6
Dhawan Lab, Institute of Liver Studies, King's College London and King's College Hospital NHS Foundation Trust, London, SE5 9PJ, UK. anil.dhawan@kcl.ac.uk.
7
Paediatric Liver, GI and Nutrition Centre, King's College Hospital Denmark Hill, London, SE5 9RS, UK. anil.dhawan@kcl.ac.uk.
8
Dhawan Lab, Institute of Liver Studies, King's College London and King's College Hospital NHS Foundation Trust, London, SE5 9PJ, UK. robin.hughes@kcl.ac.uk.

Abstract

INTRODUCTION:

Mesenchymal stem/stromal cells (MSCs) improve the metabolic function of co-cultured hepatocytes. The present study aimed to further enhance the trophic effects of co-culture with hepatocytes using hypoxic preconditioning (HPc) of the MSCs and also to investigate the underlying molecular mechanisms involved.

METHODS:

Human adipose tissue-derived MSCs were subjected to hypoxia (2 % O2; HPc) or normoxia (20 % O2) for 24 h and then co-cultured with isolated human hepatocytes. Assays of metabolic function and apoptosis were performed to investigate the hepatotrophic and anti-apoptotic effects of co-culture. Indirect co-cultures and co-culture with MSC-conditioned medium investigated the role of paracrine factors in the hepatotrophic effects of co-culture. Reactive oxygen species (ROS) activity was antagonised with N-acetylcysteine to investigate whether HPc potentiated the effects of MSCs by intracellular ROS-dependent mechanisms. Tumour necrosis factor (TNF)-α, transforming growth factor (TGF)-β1, and extracellular collagen production was determined and CASP9 and BAX/BCL-2 signalling pathways analysed to investigate the role of soluble factors, extracellular matrix deposition, and apoptosis-associated gene signalling in the effects of co-culture.

RESULTS:

HPc potentiated the hepatotrophic and anti-apoptotic effects of co-culture by ROS-dependent mechanisms. There was increased MSC TGF-β1 production, and enhanced MSC deposition of extracellular collagen, with reduced synthesis of TNF-α, as well as a downregulation of the expression of pro-apoptotic CASP9, BAX, BID and BLK genes and upregulated expression of anti-apoptotic BCL-2 in hepatocytes.

CONCLUSIONS:

HPc potentiated the trophic and anti-apoptotic effects of MSCs on hepatocytes via mechanisms including intracellular ROS, autocrine TGF-β, extracellular collagen and caspase and BAX/BCL-2 signalling pathways.

PMID:
26626568
PMCID:
PMC4667488
DOI:
10.1186/s13287-015-0218-7
[Indexed for MEDLINE]
Free PMC Article

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