BMP2 repression and optimized culture conditions promote human bone marrow-derived mesenchymal stem cell isolation

Regen Med. 2015;10(2):109-25. doi: 10.2217/rme.14.67.

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.

Keywords: BMP2; CFU-F; hypoxia; mesenchymal stem cells; physiological normoxia; transcriptome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • BMP2 protein, human
  • Bone Morphogenetic Protein 2
  • Chemokines
  • Glucose
  • Oxygen