NOOTROPIC COMPOSITION BASED ON POLYPEPTIDE COMPLEXES ISOLATED FROM GLIAL PROGENITOR CELLS UNDER CONDITIONS OF HEAT SHOCK, AND A METHOD FOR PRODUCTION THEREOF Russian patent published in 2020 - IPC C12N5/95 G01N33/48 

FIELD: biotechnology.

SUBSTANCE: invention relates to production of a nootropic composition based on polypeptide complexes recovered from glial progenitor cells in conditions of heat shock. Method involves sampling a skin biopsy material, obtaining a fibroblast culture, cryopreservation of a fibroblast culture, defrosting the fibroblast culture, adapting the fibroblast culture to xeno free cultivation conditions, reproducing the fibroblasts for producing induced pluripotent stem cells (iPSC), iPSC cultivation, iPSC cryopreservation, iPSC defrosting, iPSC differentiation into glial progenitor cells, cultivation of glial progenitor cells under heat shock conditions, collecting the conditioned medium and obtaining a protein-peptide complex. To simulate heat shock, glial precursors are cultured for hour at 40 °C in growth medium containing DMEM/F12, 2 % additive B27, 20 ng/ml FGF-2, 1 mcM purmorfamin, after which the medium is replaced with DMEM/F12 and cultivated for 6 hours, after which the conditioned medium is collected and centrifuged at 300 rpm for 5 minutes, supernatant is collected and concentrated 24 times with 3 kDa Amicon Ultra membranes, then the obtained concentrate is subjected to sterilizing filtration to obtain a nootropic composition. Composition contains proteins and polypeptides with molecular weight from 3 to 250 kDa, of which 65 % have a molecular weight in range of 10 to 60 kDa, content of heat shock proteins of not less than 40 % of total protein weight, brain-derived neurotrophic factor (BDNF) of not less than 140 pg/ml and glial cell-derived neurotrophic factor (GDNF) of not less than 60 pg/ml, vascular endothelium growth factor (VEGF) of not less than 80 pg/ml, total protein weight of not less than 1 mg/ml of preparation, wherein the cell culture of glial progenitor cells from which the composition is obtained demonstrates the expression of astroglial markers S100b and GFAP and consists of 98±2 % S100b⁺ cells.

EFFECT: invention widens the range of equipment.

2 cl, 2 dwg, 4 ex