title: Identification of Predictive Gene Markers for Multipotent Stromal Cell Proliferation.
abstract: Multipotent stromal cells (MSCs) are known for their distinctive ability to differentiate into different cell lineages, such as adipocytes, chondrocytes, and osteocytes. They can be isolated from numerous tissue sources, including bone marrow, adipose tissue, skeletal muscle, and others. Because of their differentiation potential and secretion of growth factors, MSCs are believed to have an inherent quality of regeneration and immune suppression. Cellular expansion is necessary to obtain sufficient numbers for use; however, MSCs exhibit a reduced capacity for proliferation and differentiation after several rounds of passaging. In this study, gene markers of MSC proliferation were identified and evaluated for their ability to predict proliferative quality. Microarray data of human bone marrow-derived MSCs were correlated with two proliferation assays. A collection of 24 genes were observed to significantly correlate with both proliferation assays (|r| >0.70) for eight MSC lines at multiple passages. These 24 identified genes were then confirmed using an additional set of MSCs from eight new donors using reverse transcription quantitative polymerase chain reaction (RT-qPCR). The proliferative potential of the second set of MSCs was measured for each donor/passage for confluency fraction, fraction of EdU+ cells, and population doubling time. The second set of MSCs exhibited a greater proliferative potential at passage 4 in comparison to passage 8, which was distinguishable by 15 genes; however, only seven of the genes (BIRC5, CCNA2, CDC20, CDK1, PBK, PLK1, and SPC25) demonstrated significant correlation with MSC proliferation regardless of passage. Our analyses revealed that correlation between gene expression and proliferation was consistently reduced with the inclusion of non-MSC cell lines; therefore, this set of seven genes may be more strongly associated with MSC proliferative quality. Our results pave the way to determine the quality of an MSC population for a particular cellular therapy in lieu of an extended in vitro or in vivo assay.
authors.affiliation: 1 Division of Cellular and Gene Therapies, Tumor Vaccines and Biotechnology Branch, Center for Biologics and Evaluation Research , US Food and Drug Administration, Silver Spring, Maryland.
authors.forename: Ian H
authors.initials: IH
authors.lastname: Bellayr
authors.affiliation: 2 Division of Cellular and Gene Therapies, Cellular and Tissue Therapies Branch, Center for Biologics Evaluation and Research , US Food and Drug Administration, Silver Spring, Maryland.
authors.forename: Ross A
authors.initials: RA
authors.lastname: Marklein
authors.affiliation: 2 Division of Cellular and Gene Therapies, Cellular and Tissue Therapies Branch, Center for Biologics Evaluation and Research , US Food and Drug Administration, Silver Spring, Maryland.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Lo Surdo
authors.affiliation: 2 Division of Cellular and Gene Therapies, Cellular and Tissue Therapies Branch, Center for Biologics Evaluation and Research , US Food and Drug Administration, Silver Spring, Maryland.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Bauer
authors.affiliation: 1 Division of Cellular and Gene Therapies, Tumor Vaccines and Biotechnology Branch, Center for Biologics and Evaluation Research , US Food and Drug Administration, Silver Spring, Maryland.
authors.forename: Raj K
authors.initials: RK
authors.lastname: Puri
mesh_terms: D000328:Adult; D049109:Cell Proliferation; D005260:Female; D005819:Genetic Markers; D006801:Humans; D008297:Male; D008875:Middle Aged; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D025341:Principal Component Analysis; D060888:Real-Time Polymerase Chain Reaction; D012044:Regression Analysis; D015203:Reproducibility of Results; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D055815:Young Adult
chemical_list: D005819:Genetic Markers