Hi! This is my first time to culture MSC(from MDS patients' BM), I notice there are three kinds of MSC: 1. stretched and big,irregular in shape,with sandy granule near nucleus, most frequently seen 2. thin and long,spindle-like. 3. flat and faint in color,with spider like reticulate structure in cytoplasm. These can form extraordinarily orginaized structure can they are confluent, some expand as colonies, with adipocytes in the center. What do you think of these cell,are they typical MSC in morphology? Do the different morphology signify anything? Thanks!
Hello ,
I think thin and long,spindle-like are MSC. Rest 3 cell types are differentiated cells like osteocytes, chondrocytes and adipocytes.
To confirm about differentiated cells I PREFER HISTOCHEMICAL STAINING.
osteocytes-ALIZARIN RED
chondrocytes-Alcian blue
Adipocyte- Oil-O-red
Hello Alexa,
I think that the cells you show are "young" MSCs. The morphology depend upon the culture medium, the age in culture and also the age of the donor. Now, as recommanded by ISCT guidelines, you have to verify the plasticity, the immunophenotype and the if possible also the immunotolerogenic properties of these cells.
Dear Alexa
yes these look like all MSCs however as Miss Diana just answered to you you would need to identify them...by flowcytometry, immune fluoressence, rt pcr.....
Good luck
Alexa,
I agree with Ciro and Diana/Seema that you should characterise, but they look like MSC on the whole. There are various subpopulations of MSC so they could belong to different types. Paul Genever has been doing some interesting work on subpopulations of MSC - check here for example:
http://www.ncbi.nlm.nih.gov/pubmed/26070611
Good luck!
When you isolate cells directly from the tissues , it is alwaya a heterogenous population with a minor population of stem cells.Normally long spindle shaped cells are MSCs.You can perform cell sorting through FACS by using specific MSC surface markers.Recently, it has been suggested to distinguish MSC through synthetic miRNA switches as well.
The morphology of the cells looks typical of MSCs. The 1st figure is fully confluent. however the rest of the figures depict growing stage MSCs. the characterization of these MSCs could be done by FACs and immunolocalization. May i also know what is the source of these MSCs
Cells in all four figures are looking like mesenchymal stem cell, you can confirm by immunophenotyping CD105, CD 73 and CD 90 positive. Figure 1 is looking like 100% confluent viable cells. Cells in figure 2 and 3 are looking like growth arrested cells.
Cheers...
Swati
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