METHOD FOR PREPARING TEST OBJECT OF CHICKEN EMBYO'S CORNEAL CELLS FOR ASSESSING DRUG CYTOTOXICITY

FIELD: medicine.SUBSTANCE: invention describes a method for preparing a test object for assessing drug cytotoxicity involving the use of chicken embryo's cornea and cell technology with cornea taken from 7-14 days chicken embryos to be thereafter ground and frozen in liquid nitrogen vapours at -180°C; when assessing drug cytotoxicity, the frozen material is slowly unfrozen, transferred into a centrifuge tube and washed in 0.9% NaCl for three times; the cell elements are thereafter ground to prepare a homogenous cell suspension; stroma is deposited in a centrifuge; a supernatant containing the corneal cells is transferred in a sterile test tube, and the cells are deposited again; the supernatant is removed; the deposition containing the corneal cells is added with 0.9% NaCl 1 ml and resuspended; the prepared suspension is analysed for cytosis; the cell viability is determined by staining in DNA fluorochromes in a flow cytofluorometer; then 0.9% NaCl is used to reduce the corneal cell suspension to the concentration of 5.0*10cells in 1 ml and transferred in culture bottles and added with the analysed preparations; for control, one of the bottles is added with 0.9% NaCl; it is followed by incubation in the culture flasks in a COincubator for one day; further the cell suspensions are transferred into the centrifuge tubes, centrifuged for cell deposition, and the suspension 1 ml is sampled of each sample for cytotoxicity analyses by cell staining in DNA fluorochromes; cytotoxicity of the analysed substances is assessed in the flow cytofluorometer.EFFECT: invention is characterised by high sensitivity and specificity of the cytotoxicity analysis; objective result interpretation enables using the declared method of a wide range of the preparations.3 tbl, 1 dwg Изобретение относится к медицине и описывает способ получения тест-объекта для оценки цитотоксичности лекарственных средств, включающий использование роговицы эмбриональных цыплят и клеточной технологии, причем выделяют роговицу эмбрионов цыплят 7-14 дней гестации, которую измельчают и замораживают в парах жидкого азота до -180°С, при оценке цитотоксичности лекарственных средств замороженный материал подвергают медленной разморозке, переносят в центрифужную пробирку и трехкратно отмывают в растворе NaCl 0,9%, а затем измельчают клеточные элементы до получения гомогенной клеточной суспензии, осаждают строму в центрифуге, надосадок, содержащий клетки роговицы, переносят в стерильную пробирку и вновь осаждают