METHOD FOR NON-INVASIVE SEMI-QUANTITATIVE ASSESSMENT OF VIABILITY OF MDA-MB-231 CELLS IN MULTIORGAN MICROPHYSIOLOGICAL MODELS Russian patent published in 2021 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology. RNA is isolated from the culture medium sample using the miRNeasy Serum/Plasma Kit with the addition of 0.3 mcl of GlycoBlue Coprecipitant 15 mg/ml, cDNA libraries are prepared for microRNA sequencing using CATS Small RNA-seq, cDNA libraries are sequenced using the Illumina NextSeq500 System sequencer and analyze the sequencing results using the BCGSC microRNA Profiling Pipeline data processing algorithm, followed by the presentation of data on the microRNA representation in the culture medium in the form of a binary logarithm counts per million reads mapped (CPM), wherein hsa-miR-222-3p and hsa-miR-99b-5p act as analyzed microRNAs. If the representation of hsa-miR-222-3p microRNA is at the level of more than 15.2 CPM and hsa-miR-99b-5p is not less than 12.5 CPM, then one speaks of 0% viability (100% death) of MDA-MB-231 cells. If the level of hsa-miR-99b-5p is not less than 12.5 CPM and hsa-miR-222-3p is not less than 14.3 CPM and not more than 15.2 CPM, then we speak about the viability of 25% of MDA-MB-231 cells. If the level of hsa-miR-99b-5p is not less than 12.5 CPM and hsa-miR-222-3p is less than 14.3 CPM, then 50% of MDA-MB-231 cells are said to be viability. If the level of hsa-miR-222-3p is in the range from 12.2 to 12.5 CPM and hsa-miR-99b-5p is not more than 12.5 CPM, then we speak of 75% of viable MDA-MB-231 cells. If the presence of hsa-miR-222-3p in the culture medium is less than 12.2 CPM and hsa-miR-99b-5p is not more than 12.5 CPM, this indicates that all MDA-MB-231 cells are viable.

EFFECT: use of this method will make it possible to analyze the viability of tumor cells with successive exposure to chemotherapeutic drugs. This approach will find its application in the personalized selection of chemotherapy regimens, will reduce the requirements for the number of tumor cells required for analysis, and the cost of increasing their required number. Method can also be used for semi-quantitative assessment of the viability of MDA-MB-231 cells in multiorgan microphysiological models.

1 cl, 1 dwg, 1 tbl, 1 ex