A simplified culture and polymerase chain reaction identification assay for quality control performance testing of stem cell media products.
OBJECTIVE
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.
Boucher
Shayne
S
Life Technologies, Primary and Stem Cell Systems, Frederick, Maryland 21704,USA. shayne.boucher@invitrogen.com
Lakshmipathy
Uma
U
Vemuri
Mohan
M
D050156:Adipogenesis / Q000235:genetics*; D018929:Cell Culture Techniques; D003470:Culture Media / Q000032:analysis; D050556:Fatty Acid-Binding Proteins / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011786:Quality Control; D015854:Up-Regulation
D003470:Culture Media; C493410:FABP4 protein, human; D050556:Fatty Acid-Binding Proteins