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Preparation and Expansion Methods for Human Pluripotent Stem Cell-Derived Human Retinal Pigment Epithelial Cells

机译:人多能干细胞衍生的人视网膜色素上皮细胞的制备和扩增方法

摘要

Preparation and expansion methods for human pluripotent stem cell-derived human retinal pigment epithelial cells are provided. The preparation method includes the following steps: collecting 3D-PRE spheroids derived from human pluripotent stem cells, performing mechanical separation to remove non-RPE cells and clusters which are non-pigmented and retain a pigmented RPE cell sheet, enzymatically digesting the pigmented RPE cell sheet to obtain an RPE single cell suspension, and thereby the human pluripotent stem cell-derived human retinal pigment epithelial cells are obtained. The expansion method includes centrifuging the RPE single cell suspension and removing a supernatant, resuspending in an RPE cell medium and seeding into a cell culture container precoated with extracellular matrix to allow primary culture, and subculturing the cells after cells spread out.
机译:提供了用于人多能干细胞的人视网膜色素上皮细胞的制备和扩增方法。制备方法包括以下步骤:收集源自人多能干细胞的3D-PRE球体,进行机械分离以去除非色素的非RPE细胞和簇并保留有色素的RPE细胞片,酶消化有色素的RPE细胞薄片获得RPE单细胞悬液,从而获得人多能干细胞来源的人视网膜色素上皮细胞。扩增方法包括离心RPE单细胞悬液并除去上清液,重悬于RPE细胞培养基中,并播种到预先涂有细胞外基质的细胞培养容器中以进行初次培养,以及在细胞铺展后对细胞进行亚培养。

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