Our lab has isolated mesenchymal stem cells from tissue and we have shown differentiation into adipose, osteo and condro, but we still need to demonstrate their multipotency using phenotypic characterization. Do you have any suggestions on the best way to do this? Thanks

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Options for Identification and Characterization of Pluripotent Stem Cells

Answered by:

Dr Uma Lakshmipathy

Company: Thermo Fisher Scientific, Inc.

Job Title: Principal Scientist

Answer

As per the minimal criteria defined by ISCT in 2006, in addition to in vitro differentiation of multipotent mesenchymal stromal cells (MSC) into adipocytes, osteoblasts and chondroblasts, cells must adhere to plastic and show positive expression of CD105, CD73, CD90 and absence of CD45, CD34, CD14, CD19 and HLA-DR {Dominici et al., Cytotherapy (2006) 8, 315-317}. Phenotypic characterization can be carried out via flow cytometric analysis of antibody stained cells.

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We have been working to reduce serum in our stem cell culture while maintaining pluripotent characteristics. Our standard media contains 10% FBS and we have not been able to reduce this amount successfully without the culture suffering. We recently were able to use human serum instead at 1.25% while maintaining similar pluripotent characteristics. However, we have concerns about adopting a process that uses human serum in a potential human therapy. Do you have any recommendations on supplements that might work to help us reduce the FBS down to a similar level as what we have achieved when using human serum?

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