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A multiplex PCR technique to characterize human bone marrow derived mesenchymal stem cells
Dillip Kumar Bishi, Soma Guhathakurta,
Published in
2009
PMID: 19693443
Volume: 31
   
Issue: 12
Pages: 1843 - 1850
Abstract
Human mesenchymal stem cells (MSCs), with capacity to differentiate into adipocytes, osteoblasts and chondrocytes, offer potential for the development of novel treatments. A critical question in MSCs biology is whether this cell population possesses a relatively uniform differentiation capability or is comprised of distinct subsets of progenitors committed to differentiate in particular pathways. To quantify the changes during growth of MSCs, we analyzed the mesenchymal phenotype and differentiation ability using a multi-marker PCR with six primer sets specific for CD73, CD90, CD105, CD166, CD45 and β-actin allowing a gel-based differential detection of the PCR products. To determine degree of variability of MSCs populations in terms of proliferation, cell proliferation assays were performed on expanded MSCs up to the sixth passage. At each passage, the osteogenic and adipogenic differentiation potentials of MSCs were verified by culture in inductive media. RT-PCR and cytochemical analysis revealed that, despite the loss of multipotentiality during expansion, certain markers remain expressed, indicating that these markers are unlikely to be reflective of the MSC's true 'stem cell' nature. Our results suggest that decrease in the expression of MSCs specific markers correlates with down-regulation of proliferation ability and differentiation efficiency of MSCs. © 2009 Springer Science+Business Media B.V.
About the journal
JournalBiotechnology Letters
ISSN01415492
Open AccessNo
Concepts (49)
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    ADIPOCYTES
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    ADIPOGENIC
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    Bone marrow cells
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    CELL POPULATIONS
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    Chondrocytes
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    Critical questions
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    CYTOCHEMICAL ANALYSIS
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    DEGREE OF VARIABILITY
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    DIFFERENTIAL DETECTION
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    Down-regulation
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    HUMAN BONE MARROW DERIVED MESENCHYMAL STEM CELLS
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    Human mesenchymal stem cells
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    Mesenchymal stem cell
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    MULTIPLEX PCR
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    OSTEOGENIC
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    PCR PRODUCTS
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    PRIMER SETS
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    Stem cell
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    Bone
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    Cell proliferation
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    Flowcharting
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    Gene expression
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    Multiplexing
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    Osteoblasts
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    Stem cells
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    Cell culture
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    ACTIN
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    LEUKOCYTE ANTIGEN
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    Article
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    Biosynthesis
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    BONE MARROW CELL
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    Cell differentiation
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    Chemistry
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    Culture medium
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    Culture technique
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    Gene expression profiling
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    Gene expression regulation
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    Genetics
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    Human
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    Methodology
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    Physiology
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    Polymerase chain reaction
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    Actins
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    ANTIGENS, CD
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    Cell culture techniques
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    Culture media
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    Gene expression regulation, developmental
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    Humans
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    Mesenchymal stem cells