The use of apolipoprotein A-I as a component of serum-free nutrient medium for bone marrow cell culture
Anna Yu. Gorodetskaya, Tatyana A. Tkachenko, Alexey N. Dudarev, Igor O. Cheshenko, Ivan F. Usynin
Institute of Biochemistry of Federal Research Center for Fundamental and Translational Medicin, Novosibirsk, Russia
a.pe4enkina@yandex.ru
Keywords: аполипопротеин А-I, культура клеток, клетки костного мозга, бессывороточная питательная среда, биосинтез ДНК, apolipoprotein A-I, cell culture, bone marrow cells, serum-free culture medium, DNA synthesis
Abstract
An important step in preparation of cells for cell therapy and tissue engineering is the cultivation of cells in vitro. The aim of this work is to study the effect of human apolipoprotein A-I (apoA-I) on the functional activity of cultured bone marrow cells and to show the possibility of using this protein instead of animal fetal serum. Material and methods. Bone marrow cells were cultured in 24-well plates in RPMI-1640 medium in a CO2 incubator at a temperature of 37 °C. The rate of incorporation of [14C]-leucine into the total cell protein and [3H]-thymidine into the DNA was used as an integral indicator of cell viability during cultivation. Results and discussion. It was found that the rate of DNA synthesis in bone marrow cells in the presence of apo A-I increased compared with the control group (without apo A-I) by 55 % after 8 hours, by 523 % after 24 hours and by 219 % after 48 hours. Under these conditions the rate of protein synthesis was also increased. The results indicate that the presence of apo A-I in serum-free culture medium preserves the functional activity of cultured bone marrow cells. Considering that the regulatory effect of apo A-I is achieved at a low protein concentration in the medium (15 μg/ml), isolation of apo A-I from the patient’s own blood serum will provide a practically safe nutrient medium for culturing autologous bone marrow cells with applications in personalized cell therapy and tissue engineering.
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