Year 2014 Vol. 22 No 5

SCIENTIFIC PUBLICATIONS
EXPERIMENTAL SURGERY

I.V. MAIBORODIN1, V.A. MATVEEVA1, R.V.MASLOV 2, N.V. ONOPRIENKO3, I.V. KUZNETSOVA1, G.A. CHASTIKIN1

FLUORESCENT MACROPHAGES IN THE LYMPH NODES AFTER APPLICATION OF MULTIPOTENT MESENCHYMAL STROMAL CELLS WITH TRANSFECTED GFP GENE

SBEE HPE Institute of chemical biology and fundamental medicine SB RAS 1, Novosibirsk,
SBEE HPE Moscow State Medical-Stomatological University named after A.I. Evdokimov 2,
SBEE HPE Novosibirsk State Medical University 3,
The Russian Federation

Objectives. To determine the reactions of the regional lymph nodes, caused by implantation of the autologous multipotent mesenchymal stromal cells of the bone marrow origin (AMMSCMO) for acceleration of mandible bone defect regeneration in experiment.
Methods. The reactions of rat regional lymph nodes, caused by implantation of AMMSCBM for acceleration of bone defect regeneration of the mandible bone have been studied by the methods of fluorescent light microscopy in experiment.
Results. One week after the introduction in an injury site of the mandible bone of polyhydroxyalkanoate with adsorbed AMMSCBM with a transfected GFP gene the numerous large macrophages with a set of oval fluorescent inclusions up to 20 micrometers in diameter have been detected in the regional (submandibular) lymph nodes. In the fluorescent cells not the whole cytoplasm was glowing, but oval granules, different of sizes which reached 10 micrometers. Clusters of fluorescent cells differed in density and background color from the surrounding tissue and had a clear boundary. It gives the possibility to assume the concentration of cells with the fluorescent cytoplasm in the lymphoid nodules. On the basis of the large-sized cells of irregular shape and with the presence of inclusions and location of the lymph nodes, it was concluded that these objects are considered to macrophages, where the lysosomes with the absorbed fluorescent material are glowing. Dimensions of clusters of fluorescent cells and the number of the objects with a glow in them has increased up to 2 weeks after implantation, followed by a progressive decline in sizes and number.
Conclusion. AMMSCBM introduced by this method is partially absorbed by macrophages. At destruction of the structures created from AMMSCBM, detritus is also phagocytized by macrophages. In any case these macrophages appear in the lymphoid regional nodules, where initiation of immunity reactions against DNA and GFP protein isn't excluded.

Keywords: multipotent mesenchymal stromal cells, lymph nodes, macrophages, macrophage fluorescence
p. 526-532 of the original issue
References
  1. Chanda D, Kumar S, Ponnazhagan S.Therapeutic potential of adult bone marrow-derived mesenchymal stem cells in diseases of the skeleton. J Cell Biochem. 2010 Oct 1;111(2):24957.
  2. Clines GA.Prospects for osteoprogenitor stem cells in fracture repair and osteoporosis. Curr Opin Organ Transplant. 2010 Feb;15(1):738.
  3. Volozhin AI, Vasil'ev AIu, Mal'ginov NN, Bulanova IM, Grigor'ian AS, Kiseleva EV, Cherniaev SE, Tarasenko IV. Ispol'zovanie mezenkhimal'nykh stvolovykh kletok dlia aktivizatsii reparativnykh protsessov kostnoi tkani cheliusti v eksperimente [The use of mesenchymal stem cells to enhance the reparative processes of the jawbone in the experiment]. Stomatologiia. 2010;89(1):1014.
  4. Maiborodin I.V., Matveeva V.A., Kolesnikov I.S., Drovosekov M.N., Toder M.S., Shevela A.I. Regeneratsiia povrezhdennoi kosti nizhnei cheliusti krys posle ispol'zovaniia autologichnykh stromal'nykh stvolovykh kletok kostnomozgovogo proiskhozhdeniia, adsorbirovannykh na fibrinovom sgustke [Regeneration of damaged bone of rat mandible after using autologous bone marrow stromal stem cell origin, adsorbed on the fibrin clot]. Morfologiia. 2011;(6}:7985.
  5. Campo JJ, Aponte JJ, Nhabomba AJ, Sacarlal J, Angulo-Barturen I, Jimenez-Diaz MB, Alonso PL, Dobano C.Feasibility of flow cytometry for measurements of Plasmodium falciparum parasite burden in studies in areas of malaria endemicity by use of bidimensional assessment of YOYO-1 and autofluorescence. J Clin Microbiol. 2011 Mar;49(3):96874.
  6. Watson J. Suppressing autofluorescence of erythrocytes. Biotech. Histochem. Biotech Histochem. 2011 Jun;86(3):207
  7. Duan M, Li WC, Vlahos R, Maxwell MJ, Anderson GP, Hibbs ML.Distinct macrophage subpopulations characterize acute infection and chronic inflammatory lung disease. J Immunol. 2012 Jul 15;189(2):94655.
  8. Mitchell AJ, Pradel LC, Chasson L, Van Rooijen N, Grau GE, Hunt NH, Chimini G. Technical advance: autofluorescence as a tool for myeloid cell analysis. J Leukoc Biol. 2010 Sep;88(3):597603
  9. Lei L, Tzekov R, Tang S, Kaushal S.Accumulation and autofluorescence of phagocytized rod outer segment material in macrophages and microglial cells. Mol Vis. 2012;18:10313.
  10. Luhmann UF, Robbie S, Munro PM, Barker SE, Duran Y, Luong V, Fitzke FW, Bainbridge JW, Ali RR, MacLaren RE.The drusenlike phenotype in aging Ccl2-knockout mice is caused by an accelerated accumulation of swollen autofluorescent subretinal macrophages. Invest Ophthalmol Vis Sci. 2009 Dec;50(12):593443.
  11. Potter KA, Simon JS, Velagapudi B, Capadona JR.Reduction of autofluorescence at the microelectrode-cortical tissue interface improves antibody detection. J Neurosci Methods. 2012 Jan 15;203(1):96105.
Address for correspondence:
630090, Rossiyskaya Federatsii,
g. Novosibirsk,
pr. akad. Lavrenteva, d. 8,
Institut himicheskoy biologii
i fundamentalnoy meditsinyi SO RAN,
tel.mob.: 7-913-753-07-67,
e-mail: imai@mail.ru.
Maiborodin Igor Valentinovich
Information about the authors:
Maiborodin I.V. MD, professor, leading researcher of the stem cell laboratory of SBEE HPE Institute of chemical biology and fundamental medicine SB RAS.
Matveeva V.A. PhD, senior researcher of the stem cell laboratory of SBEE HPE Institute of chemical biology and fundamental medicine SB RAS.
Maslov R.V. A post-graduate student of the hospital periodontics chair of the stomatological faculty of SBEE HPE Moscow State Medical-Stomatological University named after A.I. Evdokimov.
Onoprienko N.V. PhD, a senior lecturer of the obstetrics and gynecology chair of SBEE HPE Novosibirsk State Medical University
Kuznetsova I.V. PhD, a researcher of the stem cell laboratory of SBEE Institute of chemical biology and fundamental medicine SB RAS.
Chastikin G.A. PhD, applicant for Doctors degree of the stem cell laboratory of SBEE Institute of chemical biology and fundamental medicine SB RAS.
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