Eurasian Journal of Biosciences

The effect of low oxygen level on pluripotency and proliferation capability in bone marrow mesenchymal stem cell culture


Background: Physiologically, mesenchymal stem cells (MSCs) require optimal preconditioning in the form of low O2 tension by 1-3% in bone marrow, 10-15% in adipose tissue, and 2-9% in almost all body tissues. On the other hand, seminiferous tubules in testes require 1-6% of O2 gradation to perform spermatogenesis. Under these conditions, low O2 level (hypoxia) is required to reinforce the microenvironment to keep forming quiescent cells. Purpose: This study aims to reveal the differences between in-vitro cultured MSCs under normoxic conditions and hypoxic conditions in cell viability, their pluripotency, and proliferative capability. Method: This study is an exploratory in-vitro laboratory study on Bone Marrow Mesenchymal Stem Cells (BMSC) cultures using hypoxic conditions. A total of 21 culture plates were divided into 3 groups. The data was analyzed using the ANOVA Multivariate test. Results: The highest average of viable cells was found in the P2 group, amounting to 89.29 ± 5.36. On the other hand, the least average of viable cells was found in the P0 group, amounting to 63.67 ± 2.56. Conclusion: The cultures with hypoxic conditions and normoxic preconditions are the best culture conditions since they produce cells that were capable of maintaining the pluripotency while still having a better capability of proliferation and viability, compared to direct hypoxic conditions.


  • Bizzarri, A., Koehler, H., Cajlakovic, M., Pasic, A., Schaupp, L., Klimant, I., & Ribitsch, V. (2006). Continuous oxygen monitoring in subcutaneous adipose tissue using microdialysis. Analytica Chimica Acta, 573, 48-56.
  • Chow, A., Lucas, D., Hidalgo, A., Méndez-Ferrer, S., Hashimoto, D., Scheiermann, C.,... & Tanaka, M. (2011). Bone marrow CD169+ macrophages promote the retention of hematopoietic stem and progenitor cells in the mesenchymal stem cell niche. Journal of Experimental Medicine, 208(2), 261-271.
  • Covello, K. L., Kehler, J., Yu, H., Gordan, J. D., Arsham, A. M., Hu, C. J.,... & Keith, B. (2006). HIF-2α regulates Oct-4: effects of hypoxia on stem cell function, embryonic development, and tumor growth. Genes & development, 20(5), 557-570.
  • Ferdiansyah. (2010) Regeneration of Massive Bone Deffect with Bovine Hydroxyapatite as Scaffold Mesenchymal Stem Cell. Universitas Airlangga;
  • Forristal, C. E., Wright, K. L., Hanley, N. A., Oreffo, R. O., & Houghton, F. D. (2010). Hypoxia inducible factors regulate pluripotency and proliferation in human embryonic stem cells cultured at reduced oxygen tensions. Reproduction (Cambridge, England), 139(1), 85.
  • Gruber, H. E., Somayaji, S., Riley, F., Hoelscher, G. L., Norton, H. J., Ingram, J., & Hanley Jr, E. N. (2012). Human adipose-derived mesenchymal stem cells: serial passaging, doubling time and cell senescence. Biotechnic & Histochemistry, 87(4), 303-311.
  • Jones, L., Goodman, S (2012). Stem Cell Therapy in Orthopaedics. AAOS.;II:55–7.
  • Kelly, F. B., & Porucznik, M. A. (2014). Applying Stem Cells to Orthopaedic Conditions. American Academy of Orthopaedic Surgeon, 12, 94-96.
  • Margiana, R., Aman, R. A., Pawitan, J. A., Jusuf, A. A., Ibrahim, N., & Wibowo, H. (2019). The effect of human umbilical cord-derived mesenchymal stem cell conditioned medium on the peripheral nerve regeneration of injured rats. Electronic Journal of General Medicine, 16(6), em171.
  • Nie, Y., Han, B. M., Liu, X. B., Yang, J. J., Wang, F., Cong, X. F., & Chen, X. (2011). Identification of MicroRNAs involved in hypoxia-and serum deprivation-induced apoptosis in mesenchymal stem cells. International journal of biological sciences, 7(6), 762.
  • Rantam, F. A., Ferdiansyah, M. N., & Purwati, A. (2009). Stem cell exploration. Methods of isolation and culture.
  • Sun, Y., & Yang, L. (2014). Generation and Genetic Analysis of Transgenic Maize (Zea Mays L.) Resistant to Herbicide Glyphosate. The International Journal of Biotechnology, 3(12), 151-159.
  • Suroto, H. (2010). Efficacy of Using Allze Freeze Dried Tendon Composites and Mesemimal Stem Cells in the Reconstruction of Flexor Tendon Defects. Universitas Airlangga;
  • Utomo, D. N., & Abdul Rantam, F. (2017). Regeneration mechanism of full thickness cartilage defect using combination of freeze dried bovine cartilage scaffold-Allogenic bone marrow mesenchymal stem cells-Platelet rich plasma composite (SMPC) implantation. In Journal of Biomimetics, Biomaterials and Biomedical Engineering (Vol. 31, pp. 70-82). Trans Tech Publications Ltd.
  • Wenger, R. H., & Katschinski, D. M. (2005, August). The hypoxic testis and post-meiotic expression of PAS domain proteins. In Seminars in cell & developmental biology (Vol. 16, No. 4-5, pp. 547-553). Academic Press.


This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.