Gene expression profiles of human adipose tissue-derived mesenchymal stem cells are modified by cell culture density

Previous studies conducted cell expansion ex vivo using low initial plating densities for optimal expansion and subsequent differentiation of mesenchymal stem cells (MSCs). However, MSC populations are heterogeneous and culture conditions can affect the characteristics of MSCs. In this study, differ...

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Bibliographic Details
Published in:PloS one 2014-01, Vol.9 (1), p.e83363-e83363
Main Authors: Kim, Dae Seong, Lee, Myoung Woo, Yoo, Keon Hee, Lee, Tae-Hee, Kim, Hye Jin, Jang, In Keun, Chun, Yong Hoon, Kim, Hyung Joon, Park, Seung Jo, Lee, Soo Hyun, Son, Meong Hi, Jung, Hye Lim, Sung, Ki Woong, Koo, Hong Hoe
Format: Article
Language:English
Subjects:
Adipocytes
Adipose tissue
Adipose Tissue - cytology
Adipose Tissue - metabolism
Adult
Analysis
Antigens
Biology
Body fat
Bone marrow
Cell Count
Cell culture
Cell Culture Techniques
Cell density
Cell interactions
Cell proliferation
Cells, Cultured
Cellular signal transduction
Cluster Analysis
Confluence
Cytokines
Cytokines - genetics
Density
Deoxyribonucleic acid
DNA
DNA microarrays
Female
Fibroblasts
Gene expression
Gene Expression Profiling
Gene Expression Regulation
Genes
Humans
Immunity
Immunosuppression
Medicine
Mesenchymal stem cells
Mesenchymal Stromal Cells - cytology
Mesenchyme
Multiple myeloma
Pediatrics
Plastic surgery
Polymerase chain reaction
Reproducibility of Results
Reverse transcription
Rodents
Signal transduction
Stem cells
Tissues
Transcription (Genetics)
Transcriptome
Transduction
Vascular endothelial growth factor
Young Adult
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Summary:Previous studies conducted cell expansion ex vivo using low initial plating densities for optimal expansion and subsequent differentiation of mesenchymal stem cells (MSCs). However, MSC populations are heterogeneous and culture conditions can affect the characteristics of MSCs. In this study, differences in gene expression profiles of adipose tissue (AT)-derived MSCs were examined after harvesting cells cultured at different densities. AT-MSCs from three different donors were plated at a density of 200 or 5,000 cells/cm(2). After 7 days in culture, detailed gene expression profiles were investigated using a DNA chip microarray, and subsequently validated using a reverse transcription polymerase chain reaction (RT-PCR) analysis. Gene expression profiles were influenced primarily by the level of cell confluence at harvest. In MSCs harvested at ∼90% confluence, 177 genes were up-regulated and 102 genes down-regulated relative to cells harvested at ∼50% confluence (P2). Proliferation-related genes were highly expressed in MSCs harvested at low density, while genes that were highly expressed in MSCs harvested at high density (∼90% confluent) were linked to immunity and defense, cell communication, signal transduction and cell motility. Several cytokine, chemokine and growth factor genes involved in immunosuppression, migration, and reconstitution of damaged tissues were up-regulated in MSCs harvested at high density compared with MSCs harvested at low density. These results imply that cell density at harvest is a critical factor for modulating the specific gene-expression patterns of heterogeneous MSCs.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0083363