Abstract
Background aims
With the growing use of stem cell media technologies in research and clinical settings, there has been an increased demand for validated cell-based quality control tools that can first, routinely test performance of stem cell media products, second, verify stem cell line identity, and third, demonstrate differentiation potential. As a significant amount of time and effort is required to verify these aspects separately, especially with classic functional stains that take as along as 28 days to perform, there is a need for a quick, sensitive and validated assay with short turn around time.
Methods
Culture, gene microarray and polymerase chain reaction (PCR) methodologies were utilized in the design, development and testing of a standardized performance assay for the expansion, identity and differentiation potential of human multipotent mesenchymal stromal cells (MSC).
Results
A simplified culture- and PCR-based assay was validated and transferred into a quality control setting for performance testing of human MSC under uninduced and adipogenesis-induced conditions.
Conclusions
An effective strategy has been demonstrated for identifying candidate genes, validating a gene of interest and creating an inexpensive low-technology PCR assay for distinguishing uninduced and early stage differentiating stem cells. This approach extends published criteria guidelines for routinely detecting uninduced human MSC and their differentiated progeny.
Acknowledgements
We thank Ferenc Boldog for early discussions and insight in developing and testing PCR-based assays, Chao Yan Liu for early stage screening of primer sets, Lucas Chase for helpful comments on the manuscript, and Elizabeth Brennan and Wilson Lee for technical assistance.
Declaration of interest: All authors are employed by Life Technologies and perform work-for-hire research and development studies.