The primary culture bone marrow (PCBM) cells are often used in different cytological investigations. Here we study the behavior of porcine unstimulated PCBM cells during the cultivation. DNA cytophotometry analyses revealed that immature white blood cells (WBC) such as lymphoblasts, monoblasts and myeloid cells during cultivation process reduced the DNA content in the nuclei. This resulted in displacement of the DNA histogram to the left and increased the content of diploid nuclei. This is a result of cells unblocking which are in G2 cell cycle phase. We also observed diploid pathological cells with accessory or fragmentized nuclei. However, the data with erythroid cells (EC) somehow demonstrate the opposite tendency. During the late stage of cultivation, the number of immature EC with the tetraploid DNA is increased.
References
[1]
Dexter, T.M., Allen, T.D. and Lajtha, L.G. (1977) Conditions Controlling the Proliferation of Haemopoietic Stem Cells in Vitro. Journal of Cellular Physiology, 91, 335-344. http://dx.doi.org/10.1002/jcp.1040910303
[2]
Diamandopoulos, G.T. (1994) Human Myelopoiesis in Culture of Liquid Medium Lacking Colony Stimulating Factors: Therapeutic Implications for Cancer Patients with Leukopenia. Anticancer Research, 14, 1695-1702.
[3]
Maccari, A., Donti, G.V., Rizzo, M.T., Paltriccia, R., Tabilio, A. and Donti, E. (1991) In Vitro Bone Marrow Cell Culture and Cytogenetic Analysis in a Case of Myelodysplasia. Cancer Genetics and Cytogenetics, 56, 203-207.
http://dx.doi.org/10.1016/0165-4608(91)90172-Q
[4]
Hernández, J.M., Gutiérrez, N.C., Almeida, J., García, J.L., Sánchez, M.A., Mateo, G., Ríos, A. and San Miguel, J.F. (1998) IL-4 Improves the Detection of Cytogenetic Abnormalities in Multiple Myeloma and Increases the Proportion of Clonally Abnormal Metaphases. British Journal of Haematology, 103, 163-167.
http://dx.doi.org/10.1046/j.1365-2141.1998.00951.x
[5]
Lajhta, L.G. (1952) Culture of Human Bone Marrow in Vitro. The Reversibility between Normoblastic and Megaloblastic Series of Cells. Journal of Clinical Pathology, 2, 67-85.
[6]
Garderet, L., Kobari, L., Mazurier, C., De Witte, C., Giarratana, M.C., Pérot, C., Gorin, N.C., Lapillonne, H. and Douay, L. (2010) Unimpaired Terminal Erythroid Differentiation and Preserved Enucleation Capacity in Myelodysplastic 5q(del) Clones: A Single Cell Study. Haematologica, 95, 398-405. http://dx.doi.org/10.3324/haematol.2009.012773
[7]
Giarratana, M.C., Kobari, L., Lapillonne, H., Chalmers, D., Kiger, L., Cynober, T., Marden, M.C., Wajcman, H. and Douay, L. (2005) Ex Vivo Generation of Fully Mature Human Red Blood Cells from Hematopoietic Stem Cells. Nature Biotechnology, 23, 69-74. http://dx.doi.org/10.1038/nbt1047
[8]
Skinner, A.M., Grompe, M. and Kurre, P. (2012) Intra-Hematopoietic Cell Fusion as a Source of Somatic Variation in the Hematopoietic System. Journal of Cell Science, 125, 2837-2843. http://dx.doi.org/10.1242/jcs.100123
[9]
Karalova, E.M., Sargsyan, Kh.V., Hampikian, G.K., Voskanyan, H.E., Abroyan, L.O., Avetisyan, A.S., Hakobyan, L.A., Arzumanyan, H.H., Zakaryan, H.S. and Karalyan, Z.A. (2011) Phenotypic and Cytologic Studies of Lymphoid Cells and Monocytes in Primary Culture of Porcine Bone Marrow during Infection of African Swine Fever Virus. In Vitro Cellular & Developmental Biology-Animal, 47, 200-204. http://dx.doi.org/10.1007/s11626-010-9380-5
[10]
Van Furth, R. (1970) Origin and Kinetics of Monocytes and Macrophages. Seminars in Hematology, 7, 125-141.
[11]
Weh, H.J., von Paleske, A. and Hossfeld, D.K. (1983) Disappearance of Hypotetraploid Clones after Short-Term Culture of Leukemic Cells. A Case Report. Cancer Genetics and Cytogenetics, 10, 237-240. http://dx.doi.org/10.1016/0165-4608(83)90051-1
[12]
Strychmans, P., Cronkite, E.P., Fache, J., Fliedner, T.M. and Ramos, J. (1966) Deoxyribonucleic Acid Synthesis Time of Erythropoietic and Granulopoietic Cells in Human Beings. Nature, 13, 717-720. http://dx.doi.org/10.1038/211717a0
[13]
Bond, V.P., Odartchenko, N., Cottier, H., Feinendegen, L.E. and Cronkite, E.P. (1962) The Kinetics of the More Mature Erythrocytic Precursors Studied with Tritiated Thymidine. In: Jacobson, I.O. and Doyle, M., Eds., Grune and Stratton, New York, 173-183.
[14]
Rhodes, M.M., Kopsombut, P., Bondurant, M.C., Price, J.O. and Koury, M.J. (2008) Adherence to Macrophages in Erythroblastic Islands Enhances Erythroblast Proliferation and Increases Erythrocyte Production by a Different Mechanism than Erythropoietin. Blood, 111, 1700-1708. http://dx.doi.org/10.1182/blood-2007-06-098178
[15]
Chasis, J.A. and Mohandas, N. (2008) Erythroblastic Islands: Niches for Erythropoiesis. Blood, 112, 470-478.
http://dx.doi.org/10.1182/blood-2008-03-077883
[16]
Steinman, R.M., Machtinger, B.G., Fried, J. and Cohn, Z.A. (1978) Mouse Spleen Lymphoblasts Generated in Vitro. Recovery in High Yield and Purity after Floatation in Dense Bovine Plasma Albumin Solutions. The Journal of Experimental Medicine, 147, 279-296. http://dx.doi.org/10.1084/jem.147.2.279
[17]
Steinman, R.M., Blumencranz, J.S., Machtinger, G.B., Fried, J. and Cohn, Z.A. (1978) Mouse Spleen Lymphoblasts Generated in Vitro. The Journal of Experimental Medicine, 147, 297-315. http://dx.doi.org/10.1084/jem.147.2.297
