Expression of β-III-tubulin in the neonatal adrenal cell culture: comparison of monolayer and 3D-culture
Keywords:
adrenal cell culture, monolayer culture, 3D-culture, cytospheres, spheroids, β-III-tubulin, neuroblasts
Abstract
Some morphofunctional features of newborn piglets adrenal cell cultures obtained on the surfaces with different degrees of adhesiveness have been studied. It has been shown that during cultivation on adhesive surfaces a monolayer of fibroblast-like cells is formed (monolayer culture). When cultivated on a low-adhesive surface, the cells are assembled into three-dimensional spheroids that are in a floating state (3D culture). On adhesive surfaces, attached spheroids are eventually formed on the monolayer of fibroblast-like cells. When attached or floating spheroids are transferred to an adhesive surface migration of two morphological types of cells is observed – fibroblast-like and neuroblast-like cells. Immunocytochemical staining of neuroblasts, which migrate from both types of spheroids, showed the expression of neuronal marker β-III-tubulin. The number of fibroblast-like cells migrating from floating spheroids that were transferred to adhesive conditions is inversely proportional to the duration of the preliminary 3D culture. This approach allows obtaining a "pure" neuroblasts culture, which is uncontaminated by fibroblasts.
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References
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Agley C.C., Rowlerson A.M., Velloso C.P. et al. Isolation and quantitative immunocytochemical characterization of primary myogenic cells and fibroblasts from human skeletal muscle // J. Vis. Exp. – 2015. – Vol.95. – e52049.
Ahmed S. The culture of neural stem cells // J. Cell Biochem. – 2009. – Vol.106, no 1. – P. 1–6.
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Chung K., Sicard F., Vukicevic V. et al. Isolation of neural crest derived chromaffin progenitors from adult adrenal medulla // Stem cells. – 2009. – Vol.27, no 10. – P. 2602–2613.
Dalby M.J., Riehle M.O., Johnstone H.J. et al. Nonadhesive nanotopography: fibroblast response to poly(n-butyl methacrylate)-poly(styrene) demixed surface features // J. Biomed. Mater. Re. A. – 2003. – Vol.67, no 3. – P. 1025–1032.
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Gong S., Miao Y.L., Jiao G.Z. et al. Dynamics and correlation of serum cortisol and corticosterone under different physiological or stressful conditions in mice // PLoS One. – 2015. – Vol.10, no 2. – e0117503.
Gong X., Lin C., Cheng J. et al. Generation of multicellular tumor spheroids with microwell-based agarose scaffolds for drug testing // PLoS One. – 2015. – Vol.10, no 6. – e0130348.
Hammarback J.A., Palm S.L., Furcht L.T., Letourneau P.C. Guidance of neurite outgrowth by pathways of substratum-adsorbed laminin // Journal of Neuroscience Research. – 1985. – Vol.13, no 1–2. – P. 213–220.
Hervonen A., Hervonen H., Rechardt L. Axonal growth from the primitive sympathetic elements of human fetal adrenal medulla // Experientia. – 1972. – Vol.28, no 2. – P. 178–179.
Ivascu A., Kubbies M. Rapid generation of single-tumor spheroids for high-throughput cell function and toxicity analysis // J. Biomol. Screen. – 2006. – Vol.11, no 8. – P. 922–932.
Jin Y.Q., Liu W., Hong T.H., Cao Y. Efficient Schwann cell purification by differential cell detachment using multiplex collagenase treatment // J. Neurosci. Methods. – 2008. – Vol.170, no 1. – P. 140–148.
Kaewkhaw R., Scutt A.M., Haycock J.W. Integrated culture and purification of rat Schwann cells from freshly isolated adult tissue // Nat. Protoc. – 2012. – Vol.7, no 11. – P. 1996–2004.
Kinney M.A., Hookway T.A., Wang Y., McDevitt T.C. Engineering three-dimensional stem cell morphogenesis for the development of tissue models and scalable regenerative therapeutics // Ann. Biomed. Eng. – 2014. – Vol.42, no 2. – P. 352–367.
Kisselbach L., Merges M., Bossie A., Boyd A. CD90 Expression on human primary cells and elimination of contaminating fibroblasts from cell cultures // Cytotechnology. – 2009. – Vol.59, no 1. – P. 31–44.
Knight E., Przyborski S. Advances in 3D cell culture technologies enabling tissue-like structures to be created in vitro // J. Anat. – 2015. – Vol.227, no 6. – P. 746–756.
Kulkarni G.V., McCulloch C.A. Serum deprivation induces apoptotic cell death in a subset of Balb/c 3T3 fibroblasts // Journal of Cell Science. – 1994. – Vol.107. – P. 1169–1179.
Kuzmuk K., Schook L. Pigs as a model for biomedical sciences // The Genetics of the pig, second ed. / Eds. M.F.Rothschild, A.Ruvinsky. – CAB International, 2011. – P. 426–444.
Li H., Dai Y., Shu J. et al. Spheroid cultures promote the stemness of corneal stromal cells // Tissue Cell. – 2015. – Vol.47, no 1. – P. 39–48.
Lumb R., Schwarz Q. Sympathoadrenal neural crest cells: the known, unknown and forgotten? // Dev. Growth Differ. – 2015. – Vol.57, no 2. – P. 146–157.
Morimoto Y., Hsiao A.Y., Takeuchi S. Point-, line-, and plane-shaped cellular constructs for 3D tissue assembly // Adv. Drug. Deliv. Rev. – 2015. – Vol.95. – P. 29–39.
Niapour A., Karamali F., Karbalaie K. et al. Novel method to obtain highly enriched cultures of adult rat Schwann cells // Biotechnol. Lett. – 2010. – Vol.32, no 6. – P. 781–786.
Nyberg S.L., Hardin J., Amiot B. et al. Rapid, large-scale formation of porcine hepatocyte spheroids in a novel spheroid reservoir bioartificial liver // Liver Transpl. – 2005. – Vol.11, no 8. – P. 901–910.
Park A.M., Hayakawa S., Honda E. et al. Conditioned media from lung cancer cell line A549 and PC9 inactivate pulmonary fibroblasts by regulating protein phosphorylation // Arch. Biochem. Biophys. – 2012. – Vol.518, no 2. – P. 133–141.
Pastrana E., Silva-Vargas V., Doetsch F. Eyes wide open: a critical review of sphere-formation as an assay for stem cells // Cell Stem Cell. – 2011. – Vol.8, no 5. – P. 486–498.
Pilling D., Gomer R.H. Differentiation of circulating monocytes into fibroblast-like cells // Methods Mol Biol. – 2012. – Vol.904. – P. 191–206.
Ramgolam K., Lauriol J., Lalou C. et al. Melanoma spheroids grown under neural crest cell conditions are highly plastic migratory/invasive tumor cells endowed with immunomodulator function // PLoS One. – 2011. – Vol.6, no 4. – e18784.
Santana M., Chung K., Vukicevic V. et al. Isolation, characterization, and differentiation of progenitor cells from human adult adrenal medulla // Stem Cells Transl. Med. – 2012. – Vol.1. – P. 783–791.
Saxena Sh., Wahl J., Huber-Lang M.S. et al. Generation of murine sympathoadrenergic progenitor-like cells from embryonic stem cells and postnatal adrenal glands // PLoS One. – 2013. – Vol.8, no 5. – e64454.
Sidorenko O.S., Bozhok G.A., Legach E.I., Bondarenko T.P. Morphological and functional features of newborn piglets adrenal cells during culturing // Eastern European Scientific Journal. – 2014. – No 2. – Р. 11–19.
Sincennes M.C., Wang Y.X., Rudnicki M.A. Primary mouse myoblast purification using magnetic cell separation // Methods Mol. Biol. – 2017. – Vol.1556. – P. 41–50.
Song H., David O., Clejan S. et al. Spatial composition of prostate cancer spheroids in mixed and static cultures // Tissue Eng. – 2004. – Vol.10, no 7–8. – P. 1266–1276.
Svendsen C.N., Bhattacharyya A., Tai Y.T. Neurons from stem cells: preventing an identity crisis // Nat. Rev. Neurosci. – 2001. – Vol.2, no 11. – P. 831–834.
Vizzardelli C., Potter E., Berney T. et al. Automated method for isolation of adrenal medullary chromaffin cells from neonatal porcine glands // Cell. Transpl. – 2001. – Vol.10, no 8. – P. 689–696.
Wang Q.R., Wang B.H., Huang Y.H. et al. Purification and growth of endothelial progenitor cells from murine bone marrow mononuclear cells // J. Cell Biochem. – 2008. – Vol.103, no 1. – P. 21–29.
Wei Y., Zhou J., Zheng Z. et al. An improved method for isolating Schwann cells from postnatal rat sciatic nerves // Cell Tissue Res. – 2009. – Vol.337, no 3. – P. 361–369.
Zhou H., Aziza J., Sol J. et al. Cell therapy of pain: characterization of human fetal chromaffin cells at early adrenal medulla development // Exp. Neurol. – 2006. – Vol.198, no 2. – P. 370–381.
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How to Cite
Plaksina, K., Sidorenko, O., Legach, Y., Kovalenko, I., & Bozhok, G. (1). Expression of β-III-tubulin in the neonatal adrenal cell culture: comparison of monolayer and 3D-culture. The Journal of V.N.Karazin Kharkiv National University. Series «Biology», 28, 76-86. https://doi.org/10.26565/2075-5457-2017-28-9
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CELL BIOLOGY
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