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Regen Med ; 10(2): 109-25, 2015.
Article in English | MEDLINE | ID: mdl-25835477

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% O(2), 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.


Subject(s)
Bone Marrow Cells/cytology , Bone Morphogenetic Protein 2/physiology , Cell Culture Techniques , Mesenchymal Stem Cells/cytology , Bone Marrow/pathology , Bone Morphogenetic Protein 2/metabolism , Cell Differentiation , Cell Hypoxia , Cell Proliferation , Cells, Cultured , Chemokines/metabolism , Colony-Forming Units Assay , Computational Biology/methods , Glucose/chemistry , Humans , Immunophenotyping , Microscopy, Fluorescence , Oligonucleotide Array Sequence Analysis , Osteoblasts/cytology , Oxygen/chemistry , Signal Transduction , Transcriptome , Up-Regulation
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