Энвуретцел

Rec. INN (наименование, зарегистрированное ВОЗ)

allogeneic retinal pigment epithelial (RPE) cells derived from human embryonic stem cells (hESC). The hESC are expanded in a defined, serum-free, xeno-free culture medium on plates containing mitomycin C (MitC)-inactivated human dermal fibroblast (HDF) feeder cells, seeded initially on laminin/E-cadherin and finally on a reconstituted basement membrane matrix to generate a master cell bank (MCB).
The manufacturing of the human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells consists of 4 phases. First, expansion of the MCB-derived hESCs on plates containing MitC- inactivated HDF feeder cells, coated with laminin/E-cadherin. Second, the hESCs are induced to differentiate into RPE cells in embryoid body differentiation medium (EBDM) over a time frame of 80-140 days. Third, the resulting pigmented clusters are dissociated and expanded on gelatin-coated plates, in media containing a serum replacement additive, fetal bovine serum, basic human fibroblast growth factor (bhFGF) and 2- mercaptoethanol, with the cells monitored for medium pigmentation and typical cobblestone epithelial morphology. Finally, the cells are tested for expression of RPE-specific markers (bestrophin-1 [BEST1] and/or paired box 6 [PAX6]), before cell harvest (step 4).
The final cell population is characterized by the upregulation of RPE- specific cell marker (BEST1, PAX6, microphthalmia-associated transcription factor [MITF] and RPE-65), downregulation of embryonic stem cell genes (NANOG, octamer-binding transcription factor 4 [OCT4] and SOX-2) and the capability to phagocytose fluorescence particles (≥40%)

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