METHOD OF PRODUCING INDUCED PLURIPOTENT STEM CELLS FROM FIBROBLASTS OF PATIENTS WITH HUNTINGTON'S DISEASE

摘要

FIELD: chemistry.;SUBSTANCE: described is a method of producing induced pluripotent stem cells (iPS-cells) from skin fibroblasts of a patient with Huntington's chorea, involving production and reproduction of fibroblasts, inoculating said fibroblats on pools with monolayer density 30% in a medium of DMEM+P/S+L-Glu+10% FBS+bFGF 2 ng/mcl, infection of cells when monolayer density reaches 60% with four viruses LeGO-hOct4-MOI 5, LeGO-hSox2-MOI 5, LeGO-hc-Myc-MOI 2.5, LeGO-hKlf4-MOI 5. After four days, the cells are re-inoculated on plastic 1 to 22.5 in a medium of DMEM+P/S+L-Glu+10% FBS; after one day, the medium is replaced with ESmed and cultured for 8-10 days; the medium is replaced once in two days; mechanical re-inoculation is carried out 1 to 1 on matrigel in a medium of mTeSRI; culturing is carried out until growth of columns of morphologically similar to embryonic stem cells; the clones are collected after a week. Disclosed is a cell line obtained using the described method, which is meant for treating Huntington's chorea, which has the following morpho-molecular characteristics: size of the order of 20 mcm, large nucleus, growth in compact colonies with close contact, doubling period as embryonic stem cells, considering error, normal kryotype of 46 chromosomes, expression of Oct4, SOX2, FOXD3, HESX1, Nanog, Sall4.;EFFECT: high treatment efficiency.;2 cl, 5 dwg, 2 tbl, 1 ex