In an article I just read, they are talking about the isolation of MSC from the lacrimal gland:
You, S., Isolation and propagation of mesenchymal stem cells from the lacrimal gland. PMID: 21178145
How can they be sure they are dealing with MSC? They are showing positive staining for stem-cell markers like Nestin, but not for CD73, CD90 and CD105 etc.
Sounds like the characterization was quite minimal. The minimal criteria is just that, a basis for beginning to talk about MSC from various origins. In addition to positive expression of CD73, CD90, and CD105 and negative expression of CD11b, CD14, CD34, CD45, CD79a and HLA-DR, they should also show multilineage differentiation. Most show at least osteogenic and adipogenic differentiation. A thorough evaluation will also look for chondrogenic differentation. Many papers in the last few years have also implicated additional markers like Nestin, but the ISCT does not include these yet in their minimal definition of MSCs.
It is extremely difficult to be sure you are working with true MSCs. CD markers are notoriously ineffective. For example, although the Dominici – 2006 paper mentions negative for CD34, adventitial pericyte cells have been shown to have multipotent capacity but are CD34+. Other "markers" such as CD105 are on most Mesenchymal cells including fibroblasts (see Whitney 2009, TISSUE ENGINEERING: Part A, Volume 15, Number 1, 2009).
Even the multipotency assay should technically be performed on clonal populations to be sure.
Most people now accept the heterogeneity and test in functional assays to retrospectively say they are MSCs.
As James and Martin already mentioned, a multilineage differentiation assay is the best way to prove your working with MSCs.
Our experience with flow cytometric characterization is that a lot of adherent stem cell population express somewhat the same antigens with a few exceptions. We have demonstrated that CD140b is an antigen to characterize MSCs.
I understand the minimal criteria by ISCT,but could anyone suggest a marker present in MSCs apart from fibroblasts?I have come across some fibroblast cells lines not expressing HLA-DR just like MSCs and expressing all the markers suggested by ISCT?Is CD140b not expressed by fibroblasts?
that's the problem that really blows my mind too. few years ago i've even written the paper about it https://www.researchgate.net/publication/231175009_Current_view_of_mesenchymal_stem_cells_biology_(brief_review)?ev=prf_pub.
in fact, CD73, CD90 and CD105 are not enough too.
Article Current view of mesenchymal stem cells biology (brief review)
if the origin of mesenchymal stem cells isolation is through umbilical cord blood what are the chances of getting endothelial generations ?
i processed a number of samples and there are morphological differences between the cell types i am receiving, might be it can give us the initial idea that whether the samples processed are giving us mesenchymal cells or endothelial cells, depending on which we can go for further staining, but anywhich way CD140b is better option and a negative characterization can be done using CD31 or vWF
Minimum criteria for MSC for what? We cultivate human bone marrow MSC for clinical use. In my opinion to assess the quality and purity of the culture enough the next panel: CD90-FITC / СD105-PE/СD45-PС5 /. The cells with the phenotype СD90+СD105+СD45- excellent differentiate into adipocytes, chondrocytes and osteocytes.
I think the the minimal criteria should include characterization of Stro-1 or Stro-3 along with the minimal criteria described by Dominici et al 2006.
Stro-1 is gaining popularity in sorting of true MSCs with multipotential attributes.Stro-1 positive mesenchymal progenitor cells show a better differentiation potential and also a better paracrine factor secretome compared to conventional MSCs.
Stro-1 expression can also be categorized into three groups which are high,medium and low and the multipotential propensity and paracrine ability could vary with the group of Stro-1 positive cells.
http://www.ncbi.nlm.nih.gov/pubmed/?term=Enrichment+for+STRO-1+Expression+Enhances+the+Cardiovascular+Paracrine+Activity+of+Human+Bone+Marrow-Derived+Mesenchymal+Cell+Populations+P.J.+PSALTIS%2C1%2C2%2C3+S.+PATON%2C1%2C2+F.+SEE%2C4%2C5+A.+ARTHUR%2C1%2C2+S.+MARTIN%2C2+S.+ITESCU%2C4%2C5+S.G.+WORTHLEY%2C1%2C3+S.+GRONTHOS%2C1%2C2*+AND+A.C.W.+ZANNETTINO1%2C2
For mesenchymal stem cell, the criteria are CD90+, CD73+, CD105+, negative for CD45 and their capacity to differentiate to chrondrocyte, adipocyte and oestoblast. You could have some difference due to stem cell origin such as CD34 for adipose tissue SC. Some other adult stem cell may not express these markers and may have some others differentiation capability...
Level of expression of of CD markers vary among MSCs and Fibroblasts. Significantly higher expression of CD166 and that of CD9 significantly lower in MSCs has been reported compared to fibroblasts. CD146 expressed only in MSCs... MMP expression is very low in MSCs compared to Fibroblasts....PMID: 20528146
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