Advertisement

The Isolation and Culture of Human Cord Blood-Derived Mesenchymal Stem Cells Under Low Oxygen Conditions

Part of the Methods in Molecular Biology book series (MIMB, volume 698)

Abstract

There is growing evidence that low oxygen conditions are beneficial for in vitro stem cell culturing. Mimicking the physiological oxygen tension of the placental stem cell niche in cell expansion can ­ultimately result in more robust cell expansion. Growing evidence also suggests that hypoxic preconditioning of cells may improve therapeutic outcomes. Here we describe a scalable method that enables mesenchymal stromal cell expansion from virtually every cord blood unit, including those that would normally be disqualified from banking. In addition, the cells obtained by the described method fulfill exclusively the mesenchymal stromal cell characteristics defined by the International Society for Cellular Therapy.

Key words

Low-oxygen Hypoxia Normoxia Mesenchymal stem cell 

References

  1. 1.
    Erecinska M, Silver IA. Tissue oxygen tension and brain sensitivity to hypoxia. Respir Physiol 2001;128:263–76.PubMedCrossRefGoogle Scholar
  2. 2.
    Papandreou I, Powell A, Lim AL, Denko N. Cellular reaction to hypoxia: sensing and responding to an adverse environment. Mutat Res 2005;569:87–100.PubMedCrossRefGoogle Scholar
  3. 3.
    Ivanovic Z. Hypoxia or in situ normoxia: the stem cell paradigm. J Cell Physiol 2009;219:271–5.PubMedCrossRefGoogle Scholar
  4. 4.
    Hung SC, Pochampally RR, Hsu SC, et al. Short-term exposure of multipotent stromal cells to low oxygen increases their expression of CX3CR1 and CXCR4 and their engraftment in vivo. PLoS One 2007;2:e416.PubMedCrossRefGoogle Scholar
  5. 5.
    Bell EL, Klimova TA, Eisenbart J, Schumacker PT, Chandel NS. Mitochondrial reactive oxygen species trigger hypoxia-inducible factor-dependent extension of the replicative life span during hypoxia. Mol Cell Biol 2007;27:5737–45.PubMedCrossRefGoogle Scholar
  6. 6.
    Carrancio S, Lopez-Holgado N, Sanchez-Guijo FM, et al. Optimization of mesenchymal stem cell expansion procedures by cell separation and culture conditions modification. Exp Hematol 2008;36:1014–21.PubMedCrossRefGoogle Scholar
  7. 7.
    Michelakis ED, Rebeyka I, Wu X, et al. O2 sensing in the human ductus arteriosus: regulation of voltage-gated K+ channels in smooth muscle cells by a mitochondrial redox sensor. Circ Res 2002;91:478–86.PubMedCrossRefGoogle Scholar
  8. 8.
    Bertram C, Hass R. Cellular responses to reactive oxygen species-induced DNA ­damage and aging. Biol Chem 2008;389:211–20.PubMedCrossRefGoogle Scholar
  9. 9.
    Duranteau J, Chandel NS, Kulisz A, Shao Z, Schumacker PT. Intracellular signaling by reactive oxygen species during hypoxia in cardiomyocytes. J Biol Chem 1998;273:11619–24.PubMedCrossRefGoogle Scholar
  10. 10.
    Guzy RD, Schumacker PT. Oxygen sensing by mitochondria at complex III: the paradox of increased reactive oxygen species during hypoxia. Exp Physiol 2006;91:807–19.PubMedCrossRefGoogle Scholar
  11. 11.
    Tang YL, Zhu W, Cheng M, et al. Hypoxic preconditioning enhances the benefit of cardiac progenitor cell therapy for treatment of myocardial infarction by inducing CXCR4 expression. Circ Res 2009;104:1209–16.PubMedCrossRefGoogle Scholar
  12. 12.
    Rosova I, Dao M, Capoccia B, Link D, Nolta JA. Hypoxic preconditioning results in increased motility and improved therapeutic potential of human mesenchymal stem cells. Stem Cells 2008;26:2173–82.PubMedCrossRefGoogle Scholar
  13. 13.
    Muller I, Kordowich S, Holzwarth C, et al. Animal serum-free culture conditions for isolation and expansion of multipotent mesenchymal stromal cells from human BM. Cytotherapy 2006;8:437–44.PubMedCrossRefGoogle Scholar
  14. 14.
    Krampera M, Pasini A, Rigo A, et al. HB-EGF/HER-1 signaling in bone marrow mesenchymal stem cells: inducing cell expansion and reversibly preventing multilineage differentiation. Blood 2005;106:59–66.PubMedCrossRefGoogle Scholar
  15. 15.
    Hong L, Sultana H, Paulius K, Zhang G. Steroid regulation of proliferation and ­osteogenic differentiation of bone marrow stromal cells: a gender difference. J Steroid Biochem Mol Biol 2009;114:180–5.PubMedCrossRefGoogle Scholar
  16. 16.
    Fan X, Liu T, Liu Y, Ma X, Cui Z. Optimization of primary culture condition for mesenchymal stem cells derived from umbilical cord blood with factorial design. Biotechnol Prog 2009;25:499–507.PubMedCrossRefGoogle Scholar
  17. 17.
    Chase LG, Firpo MT. Development of serum-free culture systems for human embryonic stem cells. Curr Opin Chem Biol 2007;11:367–72.PubMedCrossRefGoogle Scholar
  18. 18.
    Reddy NP, Vemuri MC, Pallu R. Isolation of stem cells from human umbilical cord blood. Methods Mol Biol 2007;407:149–63.PubMedCrossRefGoogle Scholar
  19. 19.
    Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006;8:315–7.PubMedCrossRefGoogle Scholar
  20. 20.
    Grayson WL, Zhao F, Izadpanah R, Bunnell B, Ma T. Effects of hypoxia on human mesenchymal stem cell expansion and plasticity in 3D constructs. J Cell Physiol 2006;207:331–9.PubMedCrossRefGoogle Scholar
  21. 21.
    Milner R, Hung S, Erokwu B, Dore-Duffy P, LaManna JC, del Zoppo GJ. Increased expression of fibronectin and the alpha 5 beta 1 integrin in angiogenic cerebral blood vessels of mice subject to hypobaric hypoxia. Mol Cell Neurosci 2008;38:43–52.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Finnish Red Cross Blood ServiceHelsinkiFinland

Personalised recommendations