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ISSN 2522-9028 (Print)
ISSN 2522-9036 (Online)
DOI: https://doi.org/10.15407/fz

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. 2018; 64(6): 32-39


THE OPTIMIZATION METHOD OF ISOLATION OF IMMATURE DENDRITIC CELLS

A.N. Goltsev, T.G. Dubrava, E.E. Yampolskaya, Yu.A. Gayevskaya, N.N. Babenko, N.A. Bondarovich, I.G. Kovalenko

    Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv, Ukraine
DOI: https://doi.org/10.15407/fz64.06.032


Abstract

The method of isolation of bone marrow mononuclear cells in «trasograph» gradient, enabling to obtain more than 40 % of cells, among those 28 % were CD14+-monocytes has been presented. The minimum set of phenotype markers to assess immature dendritic cells in vitro has been selected. In a comparative aspect it has been shown that the most adequate sources of obtaining of dendritic cells are mononuclear cells in contrast to total cell suspension of bone marrow. It has been found that the differentiation of myeloid progenitor cells and monocytes into immature dendritic cells was accompanied with the loss of CD14 antigen , minimization of expression rate of co-stimulatory molecule CD86 and appearance of CD11b-antigen marker, which is the feature of these cells. The results allow not only to estimate the qualitative characteristics of the obtained dendritic cells and their tolerogenic activity in the model of autoimmune diseases, but also to underline the perspectives of establishing their stocks at low temperature banks to be used in clinics.

Keywords: dendritic cells; bone marrow; mononuclear cells, monocytes; co-stimulatory molecules

References

  1. Alcántara-Hernández M, Leylek R, Wagar LE, Engleman EG, Tibor Keler T, Marinkovich MP et al. Phenotypic Mapping of Human Dendritic Cells Reveals Interindividual Variation and Tissue Specialization. 2017. Dec 19; 47(6):1037-50.
  2.   Marín E, Cuturi MC, Moreau A. Tolerogenic Dendritic Cells in Solid Organ Transplantation: Where Do We Stand? Front Immunol. 2018 CrossRef fimmu.2018.00274  
  3. Talaev VY, Plekhanova MV, Matveichev AV. Experimental models suitable for assessing the effect of components of new vaccines being developed on the differentiation of dendritic cells. The medial. 2014; 2(12): 135-53.
  4.  
  5. Sennikov SV, Kulikova EV, Knauer NY, Hantakova UN. Molecular and cellular mechanisms mediated by dendritic cells involved in the induction of tolerance. Medical immunol. 2017; 19 (4): 359-74. CrossRef  
  6. Zhou LJ, Tedder TF. Human blood dendritic cells selectively express CD83, a member of the immunoglobulin superfamily. J Immunol. 1995;154: 3821-35. PubMed
  7.  
  8. Steinman RM, Hawiger D, Nussenzweig MC. Tolerogenic dendritic cells. Annu Rev. Immunol. 2003; 21:685-711. CrossRef PubMed
  9.  
  10. Goltsev AN, Rassokha IV, Dubrava TG, Ostankova LV, Ostankov MV; Safonov VI, ZykovaAV. Experimental therapy of graft-versus-host disease by mesenchymal stromal cells grown on oxide nanocoatings. Fiziol Zh. 2016; 62 (5):1-11. CrossRef  
  11. Boyum A. A one-stage procedure for isolation of granulocytes and lymphocytes from human blood. General sedimentation properties of white blood cells in a 1g gravity. Scand J Clin Lab Investig. 1968; 97:51-76.
  12.  
  13. Dohnal A, Gra Y, Witt V, Eichstill C, Wagner D, Ul-Haq S et al. J Cell Mol Med. 2009; 13:125-35. CrossRef PubMed PubMedCentral
  14.  
  15. Schmid MA, Kingston D, Boddupalli S, Manz MG. Instructive cytokine signals in dendritic cell lineage commitment. Immunol Rev. 2010 Mar; 234(1):32-44. CrossRef PubMed
  16.   Yoo S, Ha S-J. Generation of tolerogenic dendritic cells and their thera-peutic applications. Immune Netw. 2016;16:52-60. doi:10.4110/in.2016. 16.1.52  
  17. Romani N, Reider D, Heuer M, Ebner S, Kämpgen E, Eibl B, Niederwieser D, Schuler G.Generation of mature dendritic cells from human blood. An improved method with special regard to clinical applicability. J Immunol Methods. 1996 Sep 27;196 (2):137-51. CrossRef  
  18. Justin A, Babita A. IL-4 is more effective than IL-13 for in vitro differentiation of dendritic cells from peripheral blood mononuclear cells. Int Immunol. 2005;1:312-9.
  19.  
  20. Hettihewa LM. Prolonged expression of MHC class I - peptide expression in bone marrow derived retrovirus transfected matured dendritic cells by continuous centrifugation in the presence of IL-4. Indian J Med Res. 2011; 34(5): 672-8. CrossRef PubMed PubMedCentral
  21.  
  22. Nazarkina Zh.K., Laktionov P.P. Obtaining dendritic cells for immunotherapy of cancers. Biomedical chem. 2015; 61 (1): 30-40.
  23.  
  24. Nekhaeva TL Optimization of technology and start-up of obtaining antitumor vaccines based on autologous dendritic cells: dis. ... Candidate of Medical Sciences, FGBU «Research Institute of Oncology. N.N. Petrova «of the Ministry of Health of Russia, St. Petersburg, 2014. [Russian].
  25.  
  26. Popov I, Li Mu, Zheng X, Zhang X, Ichim TE et al. Preventing autoimmune arthritis using antigen-specific immature dendritic cells: a novel tolerogenic vaccine. Arthritis Res Ther. 2006; 8(5):R141. CrossRef PubMed PubMedCentral
  27.   Madaan A, Verma R, Singh AT, Jain SK, Jaggi M. A stepwise procedure for isolation of murine bone marrow and generation of dendritic cells. J Biol Methods. 2014;1(1):e1.doi: 10.14440/jbm.2014.12 CrossRef   Yang J, Yang Y, Ren Y, Xie R, Zou H, Fan H A Mouse Model of Adoptive Immunotherapeutic Targeting of Autoimmune Arthritis Using Allo-Tolerogenic Dendritic Cells. PLoS ONE 2013; 8(10): e77729. CrossRef pone.0077729  
  28. Liu J, Cao X. Regulatory dendritic cells in autoimmunity: a comprehensive review. J. Autoimmun. 2015; 63: 1-12. CrossRef PubMed

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