I have read and done several protocols for WJ-MSC isolation but none of them work. I have isolated some cells, but never enough for them to survive
I cut and dissect 1-3 mm3 of the umbilical cord, put those in a 50 ml tube (10 mg approx) and immerse it in 10 ml of 1 mg/ml of Collagenase II (Gibco) for 1.5 to 2 h at 37°C. I don't have a shaker with T°, so I manually shake the tube every 10 minutes. After the incubation time, I centrifuge the tube at 300 g for 5 minutes, discard the supernatant and resuspend the pellet to put it in a 72 cm2 flask.
Some protocols mention a 30 min trypsin digestion after the first digestion. Others said that a cell strainer has to be used to discard the large pieces of tissue.
Anyone has a clue?
Thank you in advanced.
There are several differentiation methods for mesenchymal stem cells (MSCs) into hepatocyte-like cell. Investigators reported various hepatic differentiation protocols such as modifying culturing conditions or using various growth factors/cytokines. this literature review compared different MSCs extraction and isolation protocols from Wharton’s jelly (WJ) and explored various MSCs differentiation methods.
Various protocols have been recommended for MSCs isolated from WJ, such as enzymatic, enzymatic-explant, and explant methods. In the explant method, valuable time is wasted, but the cost and biological contaminations are reduced and the number of isolated cells is high. However, other features, such as immune phenotype and multiline-age differentiation capacity, do not differ from other methods. There are also several differentiation methods for hepatocyte-like cell including the induction of MSC by cytokines and growth factors, and the differentiation of MSC in 2- and 3-dimensional matrix (2D and 3D). Among several cytokines, hepatocyte growth factor (HGF) and fibroblast growth factor (FGF) are essential. In the early stage of the differentiation, 2D culture is useful, and in the development stage, 3D culture system with HGF and FGF cytokines are more effective in the process of differentiation. Some studies have used 3D culture system in biocompatible scaffolds, such as alginate, collagen, gelatin, and peptide-Gly-Leu-amide (PGLA).
In conclusion, Wharton’s jelly-Mesenchymal stem cells (WJ-MSCs) can be considered as an appropriate source for hepatocyte differentiation. Moreover, we introduced the explant method as the most effective protocol.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6885715/
Wharton’s jelly-Mesenchymal stem cells (WJ-MSCs) might be a suitable candidate for stem cell therapy. They have high proliferation rates, wide multipotency, and hypo-immunogenicity. There are several differentiation methods into hepatocyte-like cells, such as induction by cytokines and growth factors, and differentiation of mesenchymal stem cells (MSCs) in 2- and 3-dimensional matrix.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6885715/
Wharton`s jelly-derived mesenchymal stem cells (WJ-MSCs), have a high proliferation valency ...
https://pubmed.ncbi.nlm.nih.gov/29436792/
What's wrong with my wharton jelly derived mscs ...can anyone help
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