Hi Felipe de Lara Janz,

Which cell type you would like to differentiate your mesenchymal stem cells (MSC) into?

Because MSCs can differentiate into chondroctyes, myocytes, epithelial cells etc. And fibroblasts also can differentiate into variety of fibroblast types.

For instance MSC differentiation depends on some specific chemicals and steroids application to the culture that triggers the differentiation process. You can find lots of laboratory protocol about this from internet.

However you should define your target cell fate. I mean you may designate which cell type you want to differentiate your MSC into?

And after differentiate process protocols, you can verify your complete differentiation by checking the specific cell surface markers.

I am a bit confused with your question. Are we talking about differentiating MSC and fibroblasts into some other cell types? Then Mehmet has already given the answer. Everything will depend on what is your target cell type. We may be able to provide you further help based on that.

But the story is entirely different, if you are talking about distinguishing MSC from fibroblasts contaminants (happens in usual MSC cultures). Flow cytometry is the best technique to evaluate and provide you an answer. You may follow the cell surface markers defined by the ISCT guidelines. You may find detailed description in Dominici et al 2006, if am not mistake.

All the best!

I also interpret you question as differentiating between MSCs and fibroblasts. Short answer: very difficult. Most MSC typical features such as trilineage differentiation and immunomodulation have also been shown for fibroblasts (depending on the source). The 'classical' marker profile set forth by ISCT (Dominici et al 2006), is exactly the same for both fibroblasts and MSCs.

In raw isolates (e.g. BMMNC) you can use CD271 and CD146 to discriminate between MSCs and fibroblasts (both are positive for CD90, CD105 etc.). At least the CD146+ cells have been shown to functionally differ in constitution of the hematopoietic niche (i.e. an 'actual' stem cell characteristic).

Both these markers are quickly lost in culture however, CD271 in a matter of hours, CD146 takes a bit longer. I guess your best bet would be a combination of CD146 (MSC high) and S100A4 (fibroblast high) to discriminate..

Some literature:

http://onlinelibrary.wiley.com/enhanced/doi/10.1002/stem.1681