Abstract
Preparation of immortalized cell lines obtained from organs and tissues of farm animals is an essential area of biotechnology. The paper presents results of continuous (immortalized) cell line preparation from a primary trypsinized cell culture of an adult rabbit kidney. Cytomorphologic analysis and karyotyping were performed during the process of subcultivation in the cell culture at passages 1, 3, 24, 31, 38, 56, 66, 75, 86, 101. Dynamics of spontaneous continuous cell line formation during long-term serial passaging was examined using standard nutrient media and fetal serum. Contrary to the known cell lines of rabbit origin (Oryctolagus cuniculus L.), immortalization was not accompanied with enhanced cell production and cell size reduction. The prepared continuous cell line in its adhesive phase was up to 200 µm in size and its productivity was about 7,000 cells/cm2. Significant differences (compared to the known cell lines) in the karyotype were detected during passaging. The formed genotype was found to be near-tetrapioid when the CCL cultural properties were stabilized at passages 66–101. The known cell lines – rabbit kidney (RK-13) and rabbit cornea (SIRC) – can be characterized as pseudotriploid basing on their karyotype. This culture demonstrated low sensitivity to viruses – causative agents of rabbit diseases and sensitivity to heterologous porcine and bovine viruses.
Highlights
SUMMARY Preparation of immortalized cell lines obtained from organs and tissues of farm animals is an essential area of biotechnology.The paper presents results of continuous cell line preparation from a primary trypsinized cell culture of an adult rabbit kidney
Cytomorphologic analysis and karyotyping were performed during the process of subcultivation in the cell culture at passages 1, 3, 24, 31, 38, 56, 66, 75, 86, 101
Contrary to the known cell lines of rabbit origin (Oryctolagus cuniculus L.), immortalization was not accompanied with enhanced cell production and cell size reduction
Summary
РЕЗЮМЕ Получение иммортализованных клеточных линий, происходящих из органов и тканей сельскохозяйственных животных, является актуальным направлением биотехнологии. В статье представлены результаты получения постоянной (иммортализованной) клеточной линии из первично трипсинизированной культуры клеток почки взрослого кролика. В процессе субкультивирования проводили цитоморфологические и кариологические исследования на 1, 3, 24, 31, 38, 56, 66, 75, 86, 101-м пассажах. При проведении длительных последовательных пассажей была прослежена динамика спонтанного формирования постоянной клеточной линии с использованием стандартных питательных сред и эмбриональной сыворотки. В отличие от известных клеточных линий, произошедших от обыкновенного домашнего кролика (Oryctolagus cuniculus L.), было отмечено, что иммортализация не сопровождалась увеличением продуктивности клеток и уменьшением их размеров. При стабилизации культуральных свойств постоянной линии клеток на 66–101-м пассажах генотип сформировался как околотетраплоидный. Ключевые слова: постоянная линия клеток, первично трипсинизированная культура, иммортализация, апоптоз, теломеры. А. Получение иммортализованной культуры клеток почки кролика.
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