title: BMP2 repression and optimized culture conditions promote human bone marrow-derived mesenchymal stem cell isolation.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  We have detailed a potential milestone in the process of refinement of the BMhMSC isolation process.
authors.affiliation: Institute for Science & Technology in Medicine, Guy Hilton Research Centre, Keele University, Stoke-on-Trent, Staffordshire, ST4 7QB, UK.
authors.forename: Alasdair Gawain
authors.initials: AG
authors.lastname: Kay
authors.forename: Tina Patricia
authors.initials: TP
authors.lastname: Dale
authors.forename: Khondoker Mehedi
authors.initials: KM
authors.lastname: Akram
authors.forename: Param
authors.initials: P
authors.lastname: Mohan
authors.forename: Karen
authors.initials: K
authors.lastname: Hampson
authors.forename: Nicola
authors.initials: N
authors.lastname: Maffulli
authors.forename: Monica A
authors.initials: MA
authors.lastname: Spiteri
authors.forename: Alicia Jennifer
authors.initials: AJ
authors.lastname: El Haj
authors.forename: Nicholas Robert
authors.initials: NR
authors.lastname: Forsyth
mesh_terms: D001853:Bone Marrow / Q000473:pathology; D001854:Bone Marrow Cells / Q000166:cytology*; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism / Q000502:physiology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D015687:Cell Hypoxia; D049109:Cell Proliferation; D002478:Cells, Cultured; D018925:Chemokines / Q000378:metabolism; D003114:Colony-Forming Units Assay; D019295:Computational Biology / Q000379:methods; D005947:Glucose / Q000737:chemistry; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008856:Microscopy, Fluorescence; D020411:Oligonucleotide Array Sequence Analysis; D010006:Osteoblasts / Q000166:cytology; D010100:Oxygen / Q000737:chemistry; D015398:Signal Transduction; D059467:Transcriptome; D015854:Up-Regulation
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D018925:Chemokines; D005947:Glucose; D010100:Oxygen
keywords: BMP2; CFU-F; hypoxia; mesenchymal stem cells; physiological normoxia; transcriptome