1. BMP2 repression and optimized culture conditions promote human bone marrow-derived mesenchymal stem cell isolation
- Author
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Khondoker M. Akram, Monica A. Spiteri, Karen Hampson, Alasdair G. Kay, Tina P. Dale, Alicia J. El Haj, Param Mohan, Nicola Maffulli, and Nicholas R. Forsyth
- Subjects
Embryology ,Chemokine ,Cell ,Cell Culture Techniques ,Biomedical Engineering ,Bone Morphogenetic Protein 2 ,Bone Marrow Cells ,Q1 ,Bone morphogenetic protein 2 ,Immunophenotyping ,Colony-Forming Units Assay ,Bone Marrow ,medicine ,Humans ,CXC chemokine receptors ,Progenitor cell ,Cells, Cultured ,Cell Proliferation ,Oligonucleotide Array Sequence Analysis ,Osteoblasts ,biology ,Mesenchymal stem cell ,Computational Biology ,Cell Differentiation ,Mesenchymal Stem Cells ,equipment and supplies ,R1 ,Molecular biology ,Cell Hypoxia ,Up-Regulation ,Oxygen ,Glucose ,medicine.anatomical_structure ,Microscopy, Fluorescence ,biology.protein ,Bone marrow ,Chemokines ,Signal transduction ,Transcriptome ,Signal Transduction - Abstract
Aim: Human mesenchymal stem cells (hMSC) are multipotent progenitor cells. We propose the optimization of hMSC isolation and recovery using the application of a controlled hypoxic environment. Materials & Methods: We evaluated oxygen, glucose and serum in the recovery of hMSC from bone marrow (BMhMSC). Colony forming units-fibroblastic, cell numbers, tri-lineage differentiation, immunofluorescence and microarray were used to confirm and characterize BMhMSC. Results: In an optimized (2% O2, 4.5 g/l glucose and 5% serum) environment both colony forming units-fibroblastic (p = 0.01) and cell numbers (p = 0.0001) were enhanced over standard conditions. Transcriptional analysis identified differential expression of bone morphogenetic protein 2 (BMP2) and, putatively, chemokine (C-X-C motif) receptor 2 (CXCR2) signaling pathways. Conclusion: We have detailed a potential milestone in the process of refinement of the BMhMSC isolation process.
- Published
- 2015
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