FIELD: medicine.
SUBSTANCE: method involves increasing biomass of human embryo stem cells of the feeder-less hESKM-05 with the use of a base mTeSR medium in the bottles coated with 0.1 % gelatin. An increasing procedure is enabled with the use and daily change of a base coDMEM medium containing 10% serum substitute SR, 100 mcg/ml of kanamycin sulphate, 1 mM L-glutamine, 4 ng/ml of a base fibroblast growth factor (bFGF), 1 mM essential amino acids. A conditioned medium is produced from a culture of mice embryo neuronal cells. The increased biomass is transferred by means of 0.5 % collagenase in bottles containing a prepared collagen-chitosan matrix in the conditioned medium or in a complete nutrient medium with neuronal factor N2 added. The medium is changed every three days.
EFFECT: invention allows producing the neuronal matrix suitable for direct transplantation.
16 dwg, 1 ex
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Authors
Dates
2011-10-20—Published
2010-04-06—Filed