METHOD FOR ISOLATION OF MESENCHYMAL STEM CELLS FROM AMNIOTIC MEMBRANE OF UMBILICAL CORD, USING CELL CULTURAL MEDIUM Russian patent published in 2022 - IPC C12N5/775 

FIELD: biotechnology.

SUBSTANCE: method for isolation of a population of mesenchymal stem cells from amniotic membrane of the umbilical cord is proposed, including cultivation of umbilical cord tissue in a cultural medium containing Dulbecco’s modified eagle medium (hereinafter – DMEM), Ham’s nutrient mixture F-12 (hereinafter – F12), medium 171 (hereinafter – M171), and fetal bovine serum (hereinafter – FBS). The invention also relates to a population of mesenchymal stem cells, isolated from amniotic membrane of the umbilical cord, in which at least about 97% or more of cells of the stem cell population express each of following markers: CD73, CD90, and CD105, and do not express following markers: CD34, CD45, and HLA-DR. The invention also relates to a pharmaceutical composition of this population of mesenchymal stem cells for cell therapy; a method for the production of a cultural medium suitable for isolation of the population of mesenchymal stem cells from amniotic membrane of the umbilical cord, including mixing of following components: 250 ml of DMEM, 118 ml of M171, 118 ml of DMEM/F12, and 12.5 ml of FBS (the final concentration is 2.5%) to obtain a volume of 498.5 ml of the cultural medium. A cell cultural medium is proposed, containing: DMEM at the final concentration from about 55 to 65 % (vol./vol.), F12 at the final concentration from about 5 to 15% (vol./vol.), M171 at the final concentration from about 15 to 30% (vol./vol.), and FBS at the final concentration from about 1 to 8% (vol./vol.). The use of a cell cultural medium is proposed for isolation of mesenchymal stem cells from amniotic membrane of the umbilical cord, as well as for cultivation of mesenchymal stem cells from amniotic membrane of the umbilical cord.

EFFECT: invention provides the creation of a highly homogenous population of mesenchymal stem cells isolated from amniotic membrane of the umbilical cord.

59 cl, 12 dwg, 1 ex