Lentiviral reprogramming of human umbilical cord-derived mesenchymal stromal cells towards hepatocyte-like cells
Scope of the method
- Human health
- Basic Research
- In vitro - Ex vivo
- Human derived cells / tissues / organs
- human adult stem cells
- hepatocyte-like cells
- lentiviral reprogramming
- umbilical cord
- in vitro liver model
- Drug-induced liver injury (DILI)
- human hepatocyte-like cells
- liver enriched transcription factor
- human adult stem cells
Human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs) express several key liver-specific transcription factors as well as hepatic progenitor markers. However, they still lack the hepatocyte nuclear factors 1-alpha (HNF1a) and 4-alpha (HNF4a), indispensable for their reprogramming towards hepatocyte-like cells. This method comprises the reprogramming of hUC-MSCs towards hepatocyte-like cells through HNF1a lentiviral over-expression. Whole genome microarray analysis revealed that the expression of the nuclear receptor retinoid X receptor (RXR) gamma and the nuclear transcription factor HNF4a, in HNF1a-transduced hUC-MSCs, was significantly upregulated compared to the control conditions. This expression was even higher than found in human hepatocytes. The same was observed for the uridine 5'-diphospho-glucuronosyltransferase (UGT) 1A family. Further, a significant upregulation was observed for alpha-foetoprotein (AFP), alpha1-antitrypsin (A1-AT), the phase I biotransformation enzymes cytochrome P450 (CYP) 1A2 and CYP2A6 and the drug transporter multidrug resistance protein (MDR) 1.
- Incubator (37 ± 1°C, 90 ± 5% humidity, 5.0 ± 1% CO2/air);
- Type 2 laminar airflow HEK293T cells;
- Ultracentrifuge Water bath (37 ± 1°C);
- Inverse-phase contrast microscope;
- Colorimetric reverse transcriptase assay;
- Human hepatocytes;
- Affymetrix microarray technologies;
- Partek Genomics Suite Software.
- Still in development
Pros, cons & Future potential
- Persistent expression of HNF1a transcription factor in hUC-MSCs;
- Endogenous induction of HNF4a expression.
Genomic integration of the lentiviral vector.
Usage of a non-integrative reprogramming method e.g. mRNA transfection.
Generation of hepatocyte-like cells for the development of functional human liver-based in vitro models for pharmaco-toxicological purposes.
References, associated documents and other information
Contact personKarolien Buyl
OrganisationsVrije Universiteit Brussel (VUB)
Pharmaceutical and Pharmacological Sciences
In Vitro Toxicology and Dermato-Cosmetology