Identification of cord blood-derived mesenchymal stem/stromal cell populations with distinct growth kinetics, differentiation potentials, and gene expression profiles.
Phenotypic heterogeneity has been observed among mesenchymal stem/stromal cell (MSC) populations, but specific genes associated with this variability have not been defined. To study this question, we analyzed two distinct isogenic MSC populations isolated from umbilical cord blood (UCB1 and UCB2). The use of isogenic populations eliminated differences contributed by genetic background. We characterized these UCB MSCs for cell morphology, growth kinetics, immunophenotype, and potential for differentiation. UCB1 displayed faster growth kinetics, higher population doublings, and increased adipogenic lineage differentiation compared to UCB2. However, osteogenic differentiation was stronger for the UCB2 population. To identify MSC-specific genes and developmental genes associated with observed phenotypic differences, we performed expression analysis using Affymetrix microarrays and compared them to bone marrow (BM) MSCs. We compared UCB1, UCB2, and BM and identified distinct gene expression patterns. Selected clusters were analyzed demonstrating that genes of multiple developmental pathways, such as transforming growth factor-beta (TGF-beta) and wnt genes, and markers of early embryonic stages and mesodermal differentiation displayed significant differences among the MSC populations. In undifferentiated UCB1 cells, multiple genes were significantly up-regulated (p < 0.0001): peroxisome proliferation activated receptor gamma (PPARG), which correlated with adipogenic differentiation capacities, hepatocyte growth factor (HGF), and stromal-derived factor 1 (SDF1/CXCL12), which could both potentially contribute to the higher growth kinetics observed in UCB1 cells. Overall, the results confirmed the presence of two distinct isogenic UCB-derived cell populations, identified gene profiles useful to distinguish MSC types with different lineage differentiation potentials, and helped clarify the heterogeneity observed in these cells.
Markov
Vladimir
V
Coriell Institute for Medical Research, Camden, NJ 08103, USA.
Kusumi
Kenro
K
Tadesse
Mahlet G
MG
William
Dilusha A
DA
Hall
Dorian M
DM
Lounev
Vitali
V
Carlton
Arlene
A
Leonard
Jay
J
Cohen
Rick I
RI
Rappaport
Eric F
EF
Saitta
Biagio
B
D053687:Adult Stem Cells / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation; D005312:Fetal Blood / Q000166:cytology*; D020869:Gene Expression Profiling; D006801:Humans; D016130:Immunophenotyping; D007700:Kinetics; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D017154:Stromal Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D015854:Up-Regulation