Cell and Organ Transplantology. 2016; 4(2):211-215.
Morphometric characteristics of TGF-β1-positive cells of fetal rat brain in vitro
Liubich L. D., Lisyany M. I., Malysheva T. A., Semenova V. M., Staino L. P., Vaslovich V. V.
The State Institution “Romodanov Neurosurgery Institute, National Academy of Medical Sciences of Ukraine”, Kyiv, Ukraine
One of the directions of cell therapy being developed for brain gliomas is the use of the neurogenic stem and progenitor cells (NSCs/NPCs). There are data on the anti-tumor and immunomodulating properties of the NSCs/NPCs the mechanisms of which were not disclosed yet. One of the potential targets for tumor therapy is the transforming growth factor β (TGF-β1) which is thought to be one of the key molecules in the regulation of proliferation, differentiation and cell survival or apoptosis. In the view of available information about the possibility of TGF-β1 production by the mammalian multipotent NSCs/NPCs, the aim of this work was to study the TGF-β1-positive cells in the dynamics of cultivation of fetal brain neurogenic cells as a potential source of anti-tumor or immunomodulating effects of these cells.
Material and methods. The fetal rat brain cells on 14th (E14) day of gestation were used as the source for cultivation in standard conditions (DМЕМ + 1 % fetal bovine serum) and studied on the 2nd and 37th day by morphometry and immunocytochemistry.
Results. In the fetal rat brain cell cultures, the TGF-β1-positive cells made 22.04 ± 2.33 % and the nestin-positive cells made 49.16 ± 10.60 % of the total cells number. The morphometric parameters of TGF-β1-positive cells exceeded the corresponding values of negative cells (average values of cross-sectional areas of the cytoplasm, cross-sectional areas of the nucleus, nuclear-cytoplasmic ratio). During cultivation the relative amount of TGF-β1-positive cells was slightly decreased 15.27 ± 9.80 % (p = 0.7) and their sizes were increased. On the 37th day of cultivation the sizes of TGF-β1-positive and their nuclei were smaller in the comparison with the TGF-β1-negative cells.
Conclusions. The presence of TGF-β1 expression by part of neurogenic cells of fetal rat brain (E14) in vitro was found, which persisted throughout cultivation (~5 weeks). Significant quantitative differences of morphometric parameters of TGF-β1-positive and negative cells were detected.
Key words: fetal rat brain cell culture; TGF-β1; nestin; morphometry; immunocytochemistry
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|1. Aboody KS, Najbauer J, Metz MZ, et al. Neural Stem Cell–Mediated Enzyme/Prodrug Therapy for Glioma: Preclinical Studies. Sci Transl Med. 2013; 5(184):184-189.
|2. Bovenberg MS. Degeling MH, Tannous BA. Advances in stem cell therapy against gliomas. Trends Mol Med. 2013; 19(5):281-291.
|3. Morshed RA, Gutova M, Juliano J, et al. Analysis of glioblastoma tumor coverage by oncolytic virus-loaded neural stem cells using MRI-based tracking and histological reconstruction. Cancer Gene Therapy. 2015; 22:55-61.
|4. Shah Kh. Stem cell therapeutics for cancer. Wiley Blackwell, 2013. 304 p.
|5. Kim SU. Neural stem cell-based gene therapy for brain tumors. Stem Cell Rev. 2011; 7(1):130-40.
|6. Liu S, Yin F, Zhao M, et al. The homing and inhibiting effects of hNSCs-BMP4 on human glioma stem cells. Oncotarget. 2016; 7(14):17920-931.
|7. Namba H, Kawaji H, Yamasaki T. Use of genetically engineered stem cells for glioma therapy. Oncol Lett. 2016; 11(1):9-15.
|8. Ahmed AU, Ulasov IV, Mercer RW, et al. Maintaining and loading neural stem cells for delivery of oncolytic adenovirus to brain tumors. Methods Mol Biol. 2012; 797:97-109.
|9. Sanai N, Alvarez-Buylla A, Berger MS. Neural stem cells and the origin of gliomas. N Engl J Med. 2005; 353(8):811-22.
|10. Shervington A, Lu C. Expression of multidrug resistance genes in normal and cancer stem cells. Cancer Invest. 2008; 26(5):535-542.
|11. Ueda R, Fujita M, Zhu X, et al. Systemic inhibition of transforming growth factor-beta in glioma-bearing mice improves the therapeutic efficacy of glioma-associated antigen peptide vaccines. Clin Cancer Res. 2009; 15(21):6551-59.
|12. Lin B, Madan A, Yoon JG, et al. Massively parallel signature sequencing and bioinformatics analysis identifies up-regulation of TGFB1 and SOX4 in human glioblastoma. PLoS ONE. 2010; 5(4):1-12.
|13. Calone I, Souchelnytskyi S. Inhibition of TGFβ signaling and its implications in anticancer treatments. Exp Oncol. 2012; 34(1):9-16.
|14. Kaminska B, Kocyk M, Kijewska M. //TGF beta signaling and its role in glioma pathogenesis. Adv Exp Med Biol. 2013; 986:171-87.
|15. Zhang J, Yang W, Zhao D, et al. Correlation between TSP-1, TGF-β and PPAR-γ expression levels and glioma microvascular density. Oncol Lett. 2014; 7(1):95-100.
|16. Frei K, Gramatzki D, Tritschler I, et al. Transforming growth factor-β pathway activity in glioblastoma. Oncotarget. 2015; 6(8):5963-77.
|17. Dubrovska AM, Souchelnytskyi SS. Low-density microarray analysis of TGF1-dependent cell cycle regulation in human breast adenocarcinoma MCG7 cell line. Biopolymers and Cell. 2014; 30(2):107-17.
|18. Pelton RW, Saxena B, Jones M, et al. Immunohistochemical localization of TGF1, TGF2, and TGF3 in the mouse embryo: expression patterns suggest multiple roles during embryonic development. J Cell Biol. 1991; 15(4):1091-105.
|19. Lu Y, Jiang F, Zheng X, et al. TGFβ1 promotes motility and invasiveness of glioma cells through activation of ADAM17. Oncol Rep. 2011; 25(5):1329-35.
|20. Beier CP, Kumar P, Meyer K, et al. The cancer stem cell subtype determines immune infiltration of glioblastoma. Stem Cells and Dev. 2012; 21(15):2753-61.
|21. Wang L, Liu Z, Balivada S, et al. Interleukin-1β and transforming growth factor-β cooperate to induce neurosphere formation and increase tumorigenicity of adherenr LN-229 glioma cells. Stem Cell Research & Therapy. 2012; 3(5):1-16.|
|22. Mints M, Souchelnytskyi S. Impact of combinations of EGF, TGFβ, 17β-oestradiol, and inhibitors of corresponding pathways on prolipheration of breast cancer cell lines. Exp Oncol. 2014; 36(2):67-71.
|23. Crane CA, Han SJ, Barry JJ, et al. TGF downregulates the activating receptor NKG2D on NK cells and CD8+ T cells in glioma patients. Neuro-Oncology. 2009; 12(1):7-13.
|24. Klassen HJ, Imfeld KL, Kirov II, et al. Expression of cytokines by multipotent neural progenitor cells. Cytokine. 2003; 22(3-4):101-106.
|25. Staflin K, Honeth G, Kalliomaki S, et al. Neural progenitor cell lines inhibit rat tumor growth in vivo. Cancer Res. 2004; 64(15):5347-54.
|26. Liu J, Götherström C, Forsberg M, et al. Human neural stem/progenitor cells derived from embryonic stem cells and fetal nervous system present differences in immunogenicity and immunomodulatory potentials in vitro. Stem Cell Res. 2013; 10(3):325-37.
|28. Encinas JM, Enikolopov G. Identifying and quantitating neural stem and progenitor cells in the adult brain. Methods Cell Biol. 2008; 85:243-72.
|29. Robertson MJ, Gip P, Schaffer DV. Neural stem cell engineering: directed differentiation of adult and embryonic stem cells into neurons. Front Biosci. 2008; 13:21-50.
|30. Kumar P, Naumann U, Aigner L, et al. Impaired TGF-β induced growth inhibition contributes to the increased proliferation rate of neural stem cells harboring mutant p53. / Am J Cancer Res. 2015; 5(11):3436-45.
|31. Daynac M, Pineda JR, Chicheportiche A, et al. TGFbeta lengthens the G1 phase of stem cells in aged mouse brain. Stem Cells. 2014; 32(12):3257-65.
|32. Pineda JR, Daynac M, Chicheportiche A, et al. Vascularderived TGF-beta increases in the stem cell niche and perturbs neurogenesis during aging and following irradiation in the adult mouse brain. EMBO Mol Med. 2013; 5(4):548-62.
|33. Bryukhovetskiy I, Shevchenko V. Molecular mechanisms of the effect of TGF-1 on U87 human glioblastoma cells. Oncol Lett. 2016; 12(2):1581-90.
|34. Liubich LD, Semenova VM, Stayno LP. Influence of rat progenitor neurogenic cells supernatant on glioma 101.8 cells in vitro. Biopolymers and Cell. 2015; 31(3):200-208.
|35. Liubich LD, Lisyany NI, Semenova VM, et al. Dynamics of CD133+ cells in cultures of glioma C6 and fetal rat brain under the neurogenic cells supernatant influence. Cell and Organ Transplantology. 2015; 3(2):151-54.
|36. Liubich LD, Lisyany NI. Vplyv supernatanta progenitornyh nejroklityn na cytotoksychnu funkciju limfocytiv u shhuriv z gliomoju [Effect of supernatant progenitor neural cells on the cytotoxic function of lymphocytes in rats with glioma]. Fiziol zhurn. – Journal of Physiology. 2015; 61(4):70-77.|
Liubich LD, Lisyany MI, Malysheva T A, Semenova VM, Staino LP, Vaslovich VV. Morphometric characteristics of TGF-β1-positive cells of fetal rat brain in vitro. Cell and Organ Transplantology. 2016; 4(2):211-215. doi:10.22494/cot.v4i2.58