Can DMEM/F12 (With 3151mg/L Dextrose)+ bFGF + L-glutamine + 10% FBS + 1% anti anti, medium differentiate umbilical cord derived MSC into differentcell lineages?
What is 1% anti anti.
bFGF would be able to force towards fibroblast differentiation. You may want to try without it. These cells are relatively sturdy to culture in basal medium.
What are these cells used for following culture?, which would also determine how to culture them.
Thank you Yagna for your response. 1% anti anti is the antimycotic + antibiotic solution we add to the medium. We have tried culturing cells without bFGF also, but it does not seem to be giving a proper cell number. We are using umbilical cord tissue explant method and umbilical cord blood mononuclear cells to isolate MSCs.
UCB derived cells seems to be changing their morphology from fibroblast like cells into irregular cells, which I suspect they are differentiating in to macrophages. Hope you could help out with our concern.
Thank you
Hhahhaha, you got me on that one!
Yeah! I do hear this from people who are isolating by plastic attachment but not from those isolating by selective enrichment.
Follow the link below for some useful info, if you had this already, keep refining your enrichment n culture techniques.
https://www.stemcell.com/critical-parameters-for-the-isolation-of-mesenchymal-stem-cells-from-umbilical-cord-blood.html
Hi Vindya, what is the substrate that your cells are adhering to?
The surface on which the cells adhere to can influence their fate:
https://www.ncbi.nlm.nih.gov/pubmed/23672518
https://www.ncbi.nlm.nih.gov/pubmed/18563822
http://pubs.acs.org/doi/abs/10.1021/am4046316
http://onlinelibrary.wiley.com/doi/10.1634/stemcells.2007-0197/pdf
Hello Dr. Stefansson,
Thank you for your response to my question . Cells are getting adhered to surface of the tissue culture flask which is plastic. We are not using any specific substrate since many researchers have obtained stromal cells using plastic adherence ability. It seems to be working, but we are a bit confused, since the cells in our cultures seems to be bit longer than the cells which are in published microscopic images. That is why I posted this question to know whether differentiation could occur without any stimulants in the medium.
Thank you
Hi Vindya, in general when you are plating cells in 10%FBS, they will adhere to serum vitronectin, which is an RGD containing serum protein and is very effective at coating TC plastic.
There might be differences in cell morphology whether you coat cells along with the FBS-media or put the cells in after FBS-media is already on the plates.
@Yagna, are you trying hard to be an obnoxious ignorant jerk? Or does it come naturally?
Dear
For another informations check some links: https://www.stemcell.com/mesencult-msc-basal-medium-human.html
Regards
Marius
Hi Vindya,
I do not think that this complete medium composition that you are using will induce any specific differentiation. Although i suggest not using bFGF for WJ cultures as it is not really required to promote proliferation. Why are you using bFGF? Did you check for the expression of the FGF receptors on these cells?
Dear Charan,
In literature, there are articles which have reported the use of bFGF in culture to isolate MSCs from Cord Blood. They have reported better proliferation rates using bFGF, but a single article had reported neural like cell differentiation from MSCs using bFGF. Since the number of cells are low I thought of using bFGF for better proliferation.
Thank you for your info Dr. Stefansson and Marius.
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