Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Towards unravelling the kinetics of an acute myeloid leukaemia model system under oxidative and starvation stress: a comparison between two- and three-dimensional cultures

Abstract

A great challenge when conducting ex vivo studies of leukaemia is the construction of an appropriate experimental platform that would recapitulate the bone marrow (BM) environment. Such a 3D scaffold system has been previously developed in our group [1]. Additionally to the BM architectural characteristics, parameters such as oxygen and glucose concentration are crucial as their value could differ between patients as well as within the same patient at different stages of treatment, consequently affecting the resistance of leukaemia to chemotherapy. The effect of oxidative and glucose stress—at levels close to human physiologic ones—on the proliferation and metabolic evolution of an AML model system (K-562 cell line) in conventional 2D cultures as well as in 3D scaffolds were studied. We observed that the K-562 cell line can proliferate and remain alive for 2 weeks in medium with glucose close to physiological levels both in 20 and 5 % O2. We report interesting differences on the cellular response to the environmental, i.e., oxidative and/or nutritional stress stimuli in 2D and 3D. Higher adaptation to oxidative stress under non-starving conditions is observed in the 3D system. The glucose level in the medium has more impact on the cellular proliferation in the 3D compared to the 2D system. These differences can be of significant importance both when applying chemotherapy in vitro and also when constructing mathematical tools for optimisation of disease treatment.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  1. 1.

    Mortera-Blanco T, Mantalaris A, Bismarck A, Panoskaltsis N (2010) Development of a three-dimensional biomimicry of human acute myeloid leukemia ex vivo. Biomaterials 31:2243–2251

  2. 2.

    Bonnet D, Dick J (1997) Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nature Med 3(7):730–737

  3. 3.

    Grech G, Avellino R, Schembri-Wismayer P (2009) Molecular mechanisms in haematological malignancies. Malta Med J 21(3):6–11

  4. 4.

    Lowenberg B, Downing J, Burnett A (1999) Acute myeloid leukemia. N Engl J Med 341(14):1051–1062

  5. 5.

    Lichtman MA, Williams JW (2001) Hematology in the aged pp 501–540. In: Beutler E, Lichtman MA, Coller BS, Kipps T, Seligsohn U (eds) Williams hematology, 6th edn. McGraw-Hill, USA

  6. 6.

    Panoskaltsis N, Reid CDL, Knight SC (2003) Quantification and cytokine production of circulating lymphoid and myeloid cells in acute myelogenous leukemia (AML). Leukemia 17:716–730

  7. 7.

    Mayani H, Flores-Figueroa E, Chavez-Gonzalez A (2009) In vitro biology of human myeloid leukaemia. Leuk Res 33:624–637

  8. 8.

    Velliou E, Fuentes-Garí M, Misener R, Pefani E, Rende M, Panoskaltsis N, Pistikopoulos EN, Mantalaris A (2014) A framework for the design, modeling and optimization of biomedical systems. Comp Aid Chem Eng 34:225–236

  9. 9.

    Collins PC, Miller WM, Papoutsakis ET (1998) Stirred culture of peripheral and cord blood hematopoietic cells offers advantages over traditional static systems for clinically relevant applications. Biotechnol Bioeng 59(5):534–543

  10. 10.

    Koller MR, Emerson SG, Palsson BO (1993) Large-scale expansion of human stem and progenitor cells from bone marrow mononuclear cells in continuous perfusion cultures. Blood 82(2):378–384

  11. 11.

    Giarratana MC, Kobari L, Lapillonen H, Chalmers D, Kiger L, Cynober T et al (2005) Ex vivo generation of fully mature human red blood cells from hematopoietic stem cells. Nat Biotechnol 23:69–74

  12. 12.

    Colmone A, Amorim M, Pontier AL, Wang S, Jablonski E, Sipkins DA (2008) Leukemic cells create bone marrow niches that disrupt the behaviour of normal hematopoietic progenitor cells. Science 19(322):1861–1865

  13. 13.

    Engelhardt M, Douville J, Behringer D, Jahne A, Smith A, Henschler A, Lange W (2001) Hematopoietic recovery of ex vivo perfusion culture expanded bone marrow and unexpanded peripheral blood progenitors after myeloblastive chemotherapy. Bone Marrow Transplant 27(3):249–259

  14. 14.

    Kirito K, Fox N, Kaushansky K (2003) Thrombopoietin stimulates Hoxb4 expression: an explanation for the favourable effects of TPO on hematopoietic stem cells. Blood 102(9):3172–3178

  15. 15.

    Levac K, Karabu F, Bhatia M (2005) Identification of growth factor conditions that reduce ex vivo cord blood progenitor expansion but not alter human repopulating function in vivo. Haematologica 90(2):166–172

  16. 16.

    Maltman DJ, Przyborski A (2010) Developments in three-dimensional cell culture technology aimed at improving the accuracy of in vitro analyses. Biochem Soc Trans 38(4):1072–1075

  17. 17.

    Panoskaltsis N, Mantalaris A, Wu DJH (2005) Engineering a mimicry of bone marrow tissue ex vivo. J Biosci Bioeng 100:28–35

  18. 18.

    Rabinowitz J, Petros W, Stuart A, Peter W (1993) Characterization of endogenous cytokine concentrations after high-dose chemotherapy with autologous bone marrow support. Blood 81(9):2452–2459

  19. 19.

    Vunjak-Novakovic G, Scadden DT (2011) Biomimetric platforms for human stem cell research. Cell Stem Cell 8:252–261

  20. 20.

    Cabrita GJM, Ferreira BS, da Silva CL, Gonçalves R, Almeida-Porada G, Cabral JMS (2003) Hematopoietic stem cells: from the bone to the bioreactor. Trends Biotechnol 21(5):233–240

  21. 21.

    Rodriguez CA, Fernandes TG, Diogo MM, da Silva CL, Cabral JM (2011) Stem cell cultivation in bioreactors. Biotechnol Adv 29(6):815–829

  22. 22.

    Lee-Thedieck C, Spatz JP (2012) Artificial niches: biomimetic materials for haematopoietic stem cell culture. Macromol Rapid Commun 33:1432–1438

  23. 23.

    Tun T, Miyoshi H, Aung T, Takahashi S, Shimizu S, Kuroha T, Yamamoto M, Ohshima N (2002) Effect of growth factors on ex vivo bone marrow cell expansion using three-dimensional matrix support. Artif Organs 26(4):333–339

  24. 24.

    Da Silva CL, Gonçalves R, Crapnell KB, Cabral JMS, Zanjani ED, Almeida-Porada G (2005) A human stromal-based serum-free culture system supports the ex vivo expansion/maintenance of bone marrow and cord blood haematopoietic stem/progenitor cell. Exp Hematol 33(7):828–835

  25. 25.

    Leisten L, Kramann R, Ventura Ferreira MS, Bovi M, Neuss S, Ziegler P, Wagner W, Knuchel R, Schneider RK (2012) 3D- co-culture of hematopoietic stem and progenitor cells and mesenchymal stem cells in collagen scaffolds as a model of the hematopoietic niche. Biomaterials 22:1736–1747

  26. 26.

    Alijtawi OS, Li D, Xiao Y, Zhang D, Ramachandran K, Stehno-Bittel L, Van Veldhuizen P, Lin TL, Kambhampati S, Garimella R (2014) A novel three-dimensional stromal-based model for in vitro chemotherapy sensitivity testing of leukemia cells. Leuk Lymphoma 55:378–391

  27. 27.

    Fecteau J-F, Messmer D, Zhang S, Cui B, Chen L, Kipps TJ (2013) Impact of oxygen concentration on growth of mesenchymal stromal cells from the marrow of patients with chronic lymphocytic leukemia. Blood 121:971–974

  28. 28.

    Wilkinson ST, Tome ME, Briehl MM (2012) Mitochondrial adaptations to oxidative stress confer resistance to apoptosis in lymphoma cells. Int J Mol Sci 13:10212–10228

  29. 29.

    Zhou FL, Zhang WG, Wei YC, Meng S, Bai GG, Bai-Wang BY, Yang HY, Tian W, Meng X, Zhang H, Chen SP (2010) Involvement of oxidative stress in the relapse of acute myeloid leukemia. J Biol Chem 285:15010–15015

  30. 30.

    Lodi A, Tiziani S, Khanim FL, Drayson MT, Gunther UL, Bunce CM, Viant MR (2011) Hypoxia Triggers Major Metabolic Changes in AML Cells without Altering Indomethacin-Induced TCA Cycle Deregulation. ACS Chem Biol 6:169–175

  31. 31.

    Herst PM, Howman RA, Neeson PJ, Berridge MV, Ritchie DS (2011) The level of glycolytic metabolism in acute myeloid leukemia blasts at diagnosis is prognostic for clinical outcome. J Leukoc Biol 89:51–55

  32. 32.

    Safinia L, Mantalaris A, Bismarck A (2005) Towards a methodology for the effective surface modification of porous polymer scaffolds. Biomaterials 26:7537–7547

  33. 33.

    Yang J, Shi G, Bei J, Wang S, Cao Y, Shang Q, Yang G, Wang W (2002) Fabrication and surface modification of macroporous poly(l-lactic acid) and poly(l-lactic-co-glycolic acid) (70/30) cell scaffolds for human skin fibroblast cell culture. J Biomed Mater Res 62:438–446

  34. 34.

    Giuntoli S, Tanturli M, Gesualdo FD, Barbetti V, Rovida E, Sbarba PD (2011) Glucose availability in hypoxia regulates the selection of chronic myeloid leukemia progenitor subsets with different resistance to imatinib-mesylate. Haematologica 96(2):204–212

  35. 35.

    Dos Santos F, Andrare PZ, Boura JS, Abecasis MM, da Silva CL, Cabral JMS (2010) Ex vivo expansion of human mesenchymal stem cells: a more effective cell proliferation kinetics and metabolism under hypoxia. J Cell Physiol 223(1):27–35

  36. 36.

    Vander Heiden MG, Cantley LC, Thompson CB (2009) Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324:1029–1033

  37. 37.

    Vander Heiden MG, Locasale JW, Swanson KD, Sharfi H, Heffron GJ, Amador-Noguez D, Christofk HR, Wagner G, Rabinowitz JD, Asara JM, Cantley LC (2010) Evidence of an alternative glycolytic pathway in rapidly proliferating cells. Science 329:1492–1499

  38. 38.

    Warburg O (1956) On the origin of cancer cells. Science 123:309–314

  39. 39.

    Barnes LM, Bentley CM, Dickson AJ (2000) Advances in animal cell recombinant protein production: GS-NS0 expression system. Cytotechnology 32:109–123

  40. 40.

    DeBerardinis RJ, Cheng T (2010) Q’s next: the diverse function of glutamine in metabolism, cell biology and cancer. Oncogene 29(3):313–324

  41. 41.

    Wise DR, Thompson CB (2010) Glutamine addiction: a new therapeutic target. Trends Biochem Sci 35:427–433

  42. 42.

    Fedt SM, Bell LM, Keibler MA, Olenchock BA, Mayers JR, Wasylenko TM, Vokes NI, Guarente L, Vander Heiden MG, Stephanopoulos G (2013) Reductive glutamine metabolism is a function of a-ketoglutarate to citrate ratio in cells. Nat Commun 4:2236

  43. 43.

    Ma CYJ, Panoskaltis N, Kumar R, Xu XY, Mantalaris A (2012) Simulation of ex vivo bone marrow culture: Application to chronic myeloid leukaemia growth model. Biochem Eng J 61:66–77

Download references

Acknowledgments

This work is supported by ERC-BioBlood (no. 340719), ERC-Mobile Project (no. 226462), by the EU 7th Framework Programme [MULTIMOD Project FP7/2007-2013, no. 238013] and by the Richard Thomas Leukaemia Research Fund. R. M. is further thankful for a Royal Academy of Engineering Research Fellowship.

Author information

Correspondence to Eirini G. Velliou.

Appendix

Appendix

Concentrations of nutrients in different growth media as detected by the NOVA bioprofiler

See Table 3.

Table 3 DMEM synthesis

Calibration curve of MTS vs live K-562 cells (2D system)

See Fig. 5.

Fig. 5
figure5

MTS standard curve of the K-562 cell line for 2D culture

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Velliou, E.G., Dos Santos, S.B., Papathanasiou, M.M. et al. Towards unravelling the kinetics of an acute myeloid leukaemia model system under oxidative and starvation stress: a comparison between two- and three-dimensional cultures. Bioprocess Biosyst Eng 38, 1589–1600 (2015). https://doi.org/10.1007/s00449-015-1401-z

Download citation

Keywords

  • Acute myeloid leukaemia
  • Starvation stress
  • Oxidative stress
  • 3D scaffolds
  • K-562 cell line