Details

Autor(en) / Beteiligte

• Litvinova, L. S.
• Shupletsova, V. V.
• Yurova, K. A.
• Khaziakhmatova, O. G.
• Gazatova, N. D.
• Todosenko, N. M.
• Khlusova, M. Yu
• Malaschenko, V. V.
• Shunkin, E. O.
• Porokhova, E. D.
• Khlusov, I. A.

Titel

Content of Nucleic Acids in the Adhesive T-Lymphoblast Jurkat Line and Their Mobility In Vitro

Ist Teil von

• Cell and tissue biology, 2021-09, Vol.15 (5), p.445-454

Ort / Verlag

Moscow: Pleiades Publishing

Erscheinungsjahr

2021

Link zum Volltext

Quelle

SpringerLink (Online service)

Beschreibungen/Notizen

• T-lymphoblast-like human leukemia cells of the Jurkat line (Jurkat T cells) form polyploid forms with a high DNA content in suspension culture. Due to pronounced genetic instability, this contributes to further transformation and development of clonal diversity (polyclonal) of the cell line. Little information is available on the adherent subpopulation of Jurkat T cells. In this work, we analyzed the content of nucleic acids in suspension (DNA) and adhesive (DNA, RNA) subpopulations of Jurkat T cells using flow cytometry and propidium iodide dye, as well as confocal laser microscopy and acridine orange dye. The morphology and mobility of large (with a diameter of more than 15 μm) Jurkat T cells adhering to plastic were studied using Cell-IQ phase-contrast microscopy in real-time. According to the intensity of fluorescence in the conditionally green wavelength range (300–530 nm: from UV to green) and conditionally red (565–800 nm: from red to far-red), three subpopulations of adherent Jurkat T cells were identified: with high, medium, and low nucleic acid content. Thus, Jurkat-T cells adhering to the plastic surface of the plates retain a pronounced heterogeneity in the DNA content characteristic of the suspension fraction, which suggests a difference in the morphofunctional properties (polyclonicity) of this subpopulation of cell culture. With a sharp increase in the total cell mass, the proportion of large (giant, 15–50 μm or more) cells attached to the plastic remained constant for 21 days of cultivation and amounted to 1% of the adhesive fraction. It was found that large Jurkat T cells (with a median diameter of 31 µm) moved along the plastic at a linear (median) speed of 38 µm/h. The polynuclear Jurkat T cells on plastic are morphologically identified, with a linear increase being revealed in the mobility of adherent cells with an increase in their diameter (regression coefficient r = 0.33, p < 0.02, n = 52). Possible cellular and molecular mechanisms of an increased number of DNA copies in some adhering Jurkat T cells are discussed. It is assumed that the discovered new property (locomotor activity) can provide polyploid (multinucleated) adhering Jurkat T cells with a significant advantage—directed migration (chemotaxis) in a growing cell population under conditions of nutrient deficiency due to a change in the nutrient medium after 3–4 days of cultivation.