Hypoxia enhances self-renewal properties and markers of mesenchymal stem cells

Main Article Content

Vivi Yustianingsih
Titiek Sumarawati
Agung Putra


Mesenchymal stem cells (MSCs) are multipotent stromal cells that express CD73, CD90, and CD105 surface markers, but not CD14, CD45, CD34, CD11b, and HLA-DR. MSCs under hypoxic conditions have the essential role of maintaining the stemness capacity by releasing several growth factors into their medium, known as hypoxia conditioned medium (HCM). This study was performed to compare the effect of percentage of HCM to normoxic medium (NM) in increasing MSC proliferation marked by proliferation rate and surface marker expression.

This study was of post-test only control group design using human umbilical cord-MSCs (hUC-MSCs) as subjects. The HCM treatment group was obtained by culturing MSCs under 5% O2, whereas the NM control group was grown under 20% O2. The hUC-MSCs were divided into 4 groups with different dose ratios of HCM to NM (25%:75%; 50%:50%; 75%:25% for P1, P2 and P3, respectively and 100% of NM for the controls). All of these groups were maintained at 37oC and the data was collected after 72 hours incubation. MSC marker expression of CD73, CD90 and CD105 was analyzed using flow cytometry and MSC proliferation by trypan blue assay.

There were significant differences in MSC marker expression of CD73, CD90 and CD105 and proliferation at all dose ratios of HCM to NM (p<0.05).

Low oxygen concentration promotes MSC proliferation and stemness thus it might be beneficial for maintaining the MSC physiologic niche in-vitro.

Article Details

How to Cite
Yustianingsih, V., Sumarawati, T., & Putra, A. (2019). Hypoxia enhances self-renewal properties and markers of mesenchymal stem cells. Universa Medicina, 38(3), 164–171. https://doi.org/10.18051/UnivMed.2019.v38.164-171
Original Articles


Kagami H, Agata H, Tojo A. Bone marrow stromal cells (bone marrow-derived multipotent mesenchymal stromal cells) for bone tissue engineering: basic science to clinical translation. Int J Biochem Cell Biol 2011;43:286-9. DOI: https://doi.org/10.1016/j.biocel.2010.12.006.

Sharma A, Rani R. Do we really need to differentiate mesenchymal stem cells into insulin-producing cells for attenuation of the autoimmune responses in type 1 diabetes: immunoprophylactic effects of precursors to insulin-producing cells. Stem Cell Res Ther 2017;8:167. doi: 10.1186/s13287-017-0615-1.

Govindasamy V, Ronald VS, Abdullah AN, et al. Differentiation of dental pulp stem cells into islet-like aggregates. J Dent Res 2011;90:646–52. doi: 10.1177/0022034510396879.

Lund P, Pilgaard L, Duroux M, et al. Effect of growth media and serum replacements on the proliferation and differentiation of adipose-derived stem cells. Cytotherapy 2009;11:189–97. doi: 10.1080/14653240902736266.

Hsieh JY, Fu YS, Chang SJ, et al. Functional module analysis reveals differential osteogenic and stemness potentials in human mesenchymal stem cells from bone marrow and Wharton’s jelly of the umbilical cord. Stem Cells Dev 2010;19:1895-910. https://doi.org/10.1089/scd.2009.0485.

Estrada JC, Albo C, Benguría A, et al. Culture of human mesenchymal stem cells at low oxygen tension improves growth and genetic stability by activating glycolysis. Cell Death Differ 2012;19:743–55. doi: 10.1038/cdd.2011.172.

Pattappa G, Thorpe SD, Jegard NC, et al. Continuous and uninterrupted oxygen tension influences the colony formation and oxidative metabolism of human mesenchymal stem cells. Tissue Eng Part C Methods 2013;19:68–79. doi: 10.1089/ten.TEC.2011.0734.

Haque N, Abu Kasim NH, Rahman MT. Optimization of pre-transplantation conditions to enhance the efficacy of mesenchymal stem cells. Int J Biol Sci. 2015;11:324–34. doi: 10.7150/ijbs.10567.

Putra A, Ridwan FB, Putridewi AI, et al. The role of TNF-á induced MSCs on suppressive inflammation by increasing TGF-â and IL-10. Open Access Maced J Med Sci 2018;6:1779–83. doi: 10.3889/oamjms.2018.404.

Ma T, Grayson WL, Fröhlich M, et al. Hypoxia and stem cell-based engineering of mesenchymal tissues. Biotechnol Prog 2009;25:32–42. doi: 10.1002/btpr.128.

Ahmed NEB, Murakami M, Kaneko S. The effects of hypoxia on the stemness properties of human dental pulp stem cells (DPSCs). Sci Rep 2016;6:35476. doi: 10.1038/srep35476.

Yamamoto Y, Fujita M, Tanaka Y, et al. Low oxygen tension enhances proliferation and maintains stemness of adipose tissue–derived stromal cells. Biores Open Access 2013;2:199–205. doi: 10.1089/biores.2013.0004.

Putra A, Pertiwi D, Milla MN, et al. Hypoxia-preconditioned MSCs have superior effect in ameliorating renal function on acute renal failure animal model. Open Access Maced J Med Sci 2019;7:305–10. doi: 10.3889/oamjms.2019.049.

An HY, Shin HS, Choi JS, et al. Adipose mesenchymal stem cell secretome modulated in hypoxia for remodeling of radiation-induced salivary gland damage. PLoS One 2015;10:1–17. doi: 10.1371/journal.pone.0141862.

Mas-Bargues C, Sanz-Ros J, Román-Domínguez A, et al. Relevance of oxygen concentration in stem cell culture for regenerative medicine. Int J Mol Sci 2019;20:1195. doi: 10.3390/ijms20051195.

Caroti CM, Ahn H, Salazar HF, et al. A novel technique for accelerated culture of murine mesenchymal stem cells that allows for sustained multipotency. Sci Rep 2017;7:13334. doi: 10.1038/s41598-017-13477-y.

Lee SC, Jeong HJ, Lee SK, et al. Hypoxic conditioned medium from human adipose-derived stem cells promotes mouse liver regeneration through JAK/STAT3 signaling. Stem Cells Transl Med 2016;5:816–25. doi: 10.5966/sctm.2015-0191.

Fotia C, Massa A, Boriani F, et al. Hypoxia enhances proliferation and stemness of human adipose-derived mesenchymal stem cells. Cytotechnology 2015;67:1073–1084. doi: 10.1007/s10616-014-9731-2.

Dengler VL, Galbraith M, Espinosa JM. Transcriptional egulation by hypoxia inducible factors. Crit Rev Biochem Mol Biol 2014;49:1–15. doi: 10.3109/10409238.2013.838205.

Shi G, Jin Y. Role of Oct4 in maintaining and regaining stem cell pluripotency. Stem Cell Res Ther 2010;1:39. doi: 10.1186/scrt39.