The balance between proliferation and transcription of angiogenic factors of mesenchymal stem cells in hypoxia.
Bridging large bone defects with mesenchymal stromal cells-seeded scaffolds remains a big challenge in orthopedic surgery, due to the lack of vascularization. Within such a cell-scaffold construct, cells are exposed to ischemic conditions. When human mesenchymal stem cells (hMSCs) encounter hypoxic conditions, they show higher cell proliferation than at ambient oxygen levels. However, when hMSCs are exposed to prolonged ischemia, cell proliferation ceases completely. Exposure of hMSCs to hypoxic conditions is known to result in the transcription of angiogenic factors (AGF), which can promote the development of new blood vessels. In this study, we investigated at which oxygen level hMSC proliferation and the transcription of AGF were optimal. Human bone marrow-derived hMSCs were cultured at 0.1, 1, 2, 3, 4, 5, and 21% oxygen. Cell proliferation over 14 days was assayed using a DNA quantification method. hMSC metabolic activity over 14 days was measured using a MTT test. Quantitative RT-PCR was used to assess mRNA levels of angiogenic factors at the tested oxygen percentages. hMSCs showed the highest cell proliferation rate at 1% oxygen. The highest corrected cell metabolic rate was found at 21% oxygen, followed by 2% oxygen. HIF1α transcription did not increase under hypoxic conditions compared to 21% oxygen conditions. However, transcription of VEGF and ANG-1 was significantly higher at 2% oxygen than at 21% O2. The optimum oxygen range at which hMSCs proliferated rapidly and angiogenic factors ANG-1 and VEGF simultaneously came to expression was from 1 to 2% oxygen.
a Department of Orthopedic Surgery , University of Groningen, University Medical Center Groningen , Groningen , The Netherlands.|b Department of Biomedical Engineering , University of Groningen, University Medical Center Groningen , Groningen , The Netherlands.
Arina T
AT
Buizer
a Department of Orthopedic Surgery , University of Groningen, University Medical Center Groningen , Groningen , The Netherlands.
Sjoerd K
SK
Bulstra
a Department of Orthopedic Surgery , University of Groningen, University Medical Center Groningen , Groningen , The Netherlands.
Albert G
AG
Veldhuizen
b Department of Biomedical Engineering , University of Groningen, University Medical Center Groningen , Groningen , The Netherlands.
Roel
R
Kuijer
D000368:Aged; D000369:Aged, 80 and over; D042683:Angiopoietin-1 / Q000096:biosynthesis*; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D015687:Cell Hypoxia; D049109:Cell Proliferation; D005260:Female; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008875:Middle Aged; D014158:Transcription, Genetic; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis*
C467630:ANGPT1 protein, human; D042683:Angiopoietin-1; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
Angiogenic factor; cell proliferation; human mesenchymal stem cells; hypoxia; oxygen