Українська English

ISSN 2522-9028 (Print)
ISSN 2522-9036 (Online)

Fiziologichnyi Zhurnal

is a scientific journal issued by the

Bogomoletz Institute of Physiology
National Academy of Sciences of Ukraine

Editor-in-chief: V.F. Sagach

The journal was founded in 1955 as
1955 – 1977 "Fiziolohichnyi zhurnal" (ISSN 0015 – 3311)
1978 – 1993 "Fiziologicheskii zhurnal" (ISSN 0201 – 8489)
1994 – 2016 "Fiziolohichnyi zhurnal" (ISSN 0201 – 8489)
2017 – "Fiziolohichnyi zhurnal" (ISSN 2522-9028)

Fiziol. Zh. 2022; 68(6): 44-50


D.I. Bilko1, I.S. Dyagil2

  1. National University of Kyiv-Mohyla Academy, Ukraine
  2. SI National Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine, Ukraine


Development of new and improvement of existing models of long-term cultivation of hematopoietic stem cells (HSC) and cord blood progenitor cells is one of the most important directions in modern cell biology, related to obtaining a sufficient amount of hematopoietic tissue for experimental and clinical use. The aim of the work was to investigate the functional activity of hematopoietic progenitor cells from cord blood during long-term cultivation on a soft polyacrylamide gel substrate in the presence of a cytokine complex and to determine the advantages of this method of maintaining hematopoiesis for further use in transplantation. To realize this purpose, the following methods were used: the method of long-term cultivation in vitro on a hydrogel substrate, the method of colony formation in semi-liquid agar, cytological research methods, light and inverted microscopy, statistical research methods. Due to the use of the hydrogel substrate together with the cytokine complex, a high proliferative activity of hematopoietic cells is observed, which is reflected in their expansion during long-term cultivation (5 weeks), as well as of high colony-forming activity (521.5 ± 7.5.105 per explanted cells). Thus, the stiffness of the substrate must be taken into account for the expansion of stem cells and their immediate descendants. The presence of a soft substrate made of polyacrylamide gel along with a complex of cytokines ensures the expansion of hematopoietic cells due to the long- term support of hematopoiesis.

Keywords: hematopoietic progenitor cells; proliferation; colony formation; hydrogel substrate; long-term culture in vitro.


  1. Bilko DI, Chaikovsky YuB. The role of substrate stiffness in maintaining pluripotecy of embryonic stem cells in vitro culture. Fiziol Zh. 2021;67(3):27-34. [Ukrainian]. CrossRef
  2. Nakahata T, Ogawa M. Hemopoietic colony forming cells in umbilical cord blood with extensive capability to gene- rate mono- and multipotential hemopoietic progenitors. Clin Invest. 1982;70:1324-8. CrossRef PubMed PubMedCentral
  3. Zhang J, Yang C, Chen J, Luo M, Qu Y, Mu D, Chen Q. Umbilical cord mesenchymal stem cells and umbilical cord blood mononuclear cells improve neonatal rat memory after hypoxia-ischemia. Behav Brain Res. 2019 Apr 19;362:56-63. CrossRef PubMed
  4. De Wynter E, Ploemacher RE. Assays for the assessment of human hematopoietic stem cells. J Biol Regul Homeost Agents. 2001;15(1):23-7.
  5. Majeti R, Park CY, Weissman IL. Identification of a hierarchy of multipotent hematopoietic progenitors in human cord blood. Cell Stem Cell. 2007;131:635-45. CrossRef PubMed PubMedCentral
  6. Bilko N, Votjakova I, Vasylovska S, Bilko D. Charac- terization of the interactions between stromal and haematopoietic progenitor cells in expansion cell culture models. Cell Biol Int. 2005 Jan;29(1):83-6. CrossRef PubMed
  7. Madlambayan GJ, Rogers I, Casper RF, Zandstra PW. Controlling culture dynamics for the expansion of he- matopoietic stem cells. J Hematother Stem Cell Res. 2001;10:481-92. CrossRef PubMed
  8. Li X, Ma R, Gu Q, Liang L, Wang L, Zhang Y, et al. A fully defined static suspension culture system for large-scale human embryonic stem cell production. Cell Death Dis. 2018 Aug 30;9:892. CrossRef PubMed PubMedCentral
  9. Kuchma MD, Kyryk VM, Svitina HM, Shablii YM, Lukash LL, Lobyntseva GS, Shablii VA. Cоmparative analysis of the hеmatopoietic progenitor cells from placenta, сord blood and fetal liver, based on their im- munophenotype. BioMed Res Int. 2015;2015:418752. CrossRef PubMed PubMedCentral
  10. Mayani H, Dragowska W, Lansdorp PM. Characterization of functionally distinct subpopulations of CD34+ cord blood cells in serum free long-term cultures supplemented with hematopoietic cytokines. Blood. 1993;82(9):2664-72. CrossRef PubMed
  11. Indumathi S, Harikrishnan R, Rajkumar JS, Dhanasekaran M. Immunophenotypic comparison of heterogenous non-sorted versus sorted mononuclear cells from human umbilical cord blood: a novel cell enrichment approach. Cytotechnology. 2013;1:107-14. CrossRef PubMed PubMedCentral
  12. Chowdhury F, Li Y, Chuin Poh Y, Tamaki T, Wang N, Tanaka T. Soft substrates promote homogeneous selfre- newal of embryonic stem cells via downregulating cell matrix tractions. PLoS One. 2010 Dec 13;5(12):15655. CrossRef PubMed PubMedCentral
  13. Gerardo H, Lima A, Carvalho J, Ramos JRD, Couceiro S, Travasso RDM, Pires das Neves R, Graos M. Soft culture substrates favor stem-like cellular phenotype and facilitate reprogramming of human mesenchymal stem/ stromal cells (hMSCs) through mechanotransduction. Sci Rep. 2019 Jun 24;9(1):9086. CrossRef PubMed PubMedCentral
  14. Congrains А, Bianco J, Rosa RG, Mancuso RI, Saad STO. 3D scaffolds to model the hematopoietic stem cell niche: Applications and perspectives. Materials (Basel, Switzerland). 2021 Jan;14(3):569. CrossRef PubMed PubMedCentral
  15. Zhang P, Zhang C, Li J, Han J, Liu X, Yang H. The physical microenvironment of hematopoietic stem cells and its emerging roles in engineering applications. Stem Cell Res Therapy. 2019 Nov 19;10(1):327. CrossRef PubMed PubMedCentral
  16. Goetzke R, Sechi A, De Laporte L, Neuss S, Wagner W. Why the impact of mechanical stimuli on stem cells remains a challenge. Cell Mol Life Sci. 2018 Sep;75(18):3297-312. CrossRef PubMed
  17. Bilko DI, inventor; Method of long-term cultivation of hematopoietic stem cells. Patent No. 146819. 2021 Mar 1.
  18. Moore MAS. Cytokine and chemokine networks influencing stem cell proliferation, differentiation, and marrow homing. J Cell Biochem Suppl. 2002;38:29-38. CrossRef PubMed
  19. Laurenti E, Göttgens B. From hematopoietic stem cells to complex differentiation landscapes. Nature. 2018;553(7689):418-26. CrossRef PubMed PubMedCentral
  20. Gluckman E. Cord blood transplantation. Biol Blood Marrow Transplant. 2006;12(8):808-12. CrossRef PubMed

© National Academy of Sciences of Ukraine, Bogomoletz Institute of Physiology, 2014-2024.