I have used a cocktail of growth factors HGF, FGF, OSM and all. But mesenchymal stem cells isolated from an umbilical cord are not going towards endodermal lineage. Please suugest suitable cocktail of growth factors for hepatic differentiation. What are the factors affecting hepatic differentiation?
MSCs from umblical cord have been differentiated into hepatocytes by overexpressing hTERT. Try using it. Here is the reference for the protocol:
Liang XJ et al. (2012) 36:215-221.
Another paper using just the FGF-4 growth factor was also published:
Zhang YN et al. (2009) Cytotherapy 11:548-558.
Other references:
Campard D et al. (2008) Gastroenterology 134:833-848.
Zhao et al (2009) Cytotherapy 11:414-426.
Thanks raj. but i have used all the combinations of growth factors mentioned in these papers but cells are not getting differentiated :(
Hi Swati,
This is the basic cocktail:-
Iscove’s
Modified Dulbecco’s Medium (IMDM; GIBCO, CA)
supplemented with 20 ng/ml hepatocyte growth factor
(HGF), 10 ng/ml fibroblast growth factor-4 (FGF-4),
10 ng/ml oncostain M (OSM) (Peprotech, UK),
40 μg/L dexamethasone (DEX; Sigma-Aldrich, MO),
and 1× ITS+ Premix (BD Biosciences, CA).
What are the specifications you are looking at post induction?Are you looking for Albumin,AFP and cytokeratin 18/19??What is the change in morphology like?maybe you should play around with the concentrations of HGF and FGF-4.Check if your growth factors are active.
Regards
Charan
Stempeutics Research
Dear Charan,
Why IMDM is more preferred over LG-DMEM for hepatic differentiation? Why cant we use LG-DMEM?
Dear Akshata,
IMDM is a better modified and maybe standardized for hepatocyte differentiation.It contains selenium and more amino acids and vitamins than DMEM.It also lacks ferric nitrate which is replaced with potassium nitrate.
Regards
Charan
I want to ask that all the umbilical cord derived mesenchymal stem cells (cd 44, cd73, cd90, cd105 positive ) can go in hepatic lineage ? @ charan thej
Dear Swathi,
I strongly believe that all the UC-MSCs may not differentiate into the cells of the endodermal lineage/hepatocyte lineage efficiently.Only a certain percentage of cells differentiate into hepatocyte like cells where you can see positivity for AFP and albumin and maybe you will be able to detect secreted albumin by ELISA.
It is always recommended to use a positive liver cell line in all your experiments for comparision,maybe chang liver or other cell lines.
What is your goal here?Do you think that transplanted UC-MSC/WJ-MSCs can differentiate into hepatocytes in-vivo??
Regards
Charan
my aim is only hepatic differentiation and albumin detection in late stage and AFP detection in early stage, EROD ASSAY also if it will be mature one.
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