产品号 #100-1365_C
无血清、无异种成分培养基，用于生成功能齐全、成熟的 hPSC 衍生视网膜色素上皮
总览
无需手动筛选或细胞富集，即可快速成熟高纯度、功能性人多能干细胞 (hPSC) 衍生的视网膜色素上皮 (RPE) 细胞。STEMdiff™-XF RPE 成熟培养基是一种无异种成分、无血清培养基，支持在 35 天内将未成熟 RPE 分化为成熟且具有功能的 RPE。使用 STEMdiff™-XF RPE 成熟培养基生成的成熟 RPE 细胞表达成熟标志物（图 2），并展现出与原代 RPE 细胞相当的关键功能，包括色素沉着、多边形形态、极化和吞噬功能。本产品经过优化，可用于使用STEMdiff™-ACF RPE 分化试剂盒生成的 iPSC 衍生 RPE 细胞的成熟过程。STEMdiff™-XF RPE 成熟培养基 可与以下兼容的 STEMdiff™ RPE 产品组成一个完整且优化的工作流程：
STEMdiff™-ACF RPE 铺板补充剂
STEMdiff™-ACF RPE 分化试剂盒
如您计划将本产品用于商业或临床用途，请与我们联系。
细胞类型
多能干细胞
应用
细胞培养
品牌
STEMdiff
研究领域
疾病建模，药物发现和毒理检测，神经科学，细胞治疗开发
Data Figures
Figure 1. Workflow for the Differentiation of hPSCs into Retinal Pigment Epithelium (RPE) with STEMdiff™ RPE Culture System
hPSC colonies, previously harvested and seeded as clumps, are put directly into the medium provided in STEMdiff™-ACF RPE Differentiation Kit. Seeding into STEMdiff™ RPE Differentiation Medium A induces cells toward immature RPE. A full medium change is performed on Day 1 with fresh STEMdiff™ RPE Differentiation Medium A and then on Day 2 with STEMdiff™ RPE Differentiation Medium B. On Days 4 and 6, medium changes are performed with fresh STEMdiff™ RPE Differentiation Medium C. On Days 7 and every second day thereafter a medium change is performed with fresh STEMdiff™ RPE Differentiation Medium D. On Day 14, immature RPE cells are enzymatically harvested and subcultured in STEMdiff™-XF RPE Maturation Medium with the addition of STEMdiff™-ACF RPE Plating Supplement from Days 14 to 21 to improve survival after passaging. RPE cells begin to mature over the course of 5 weeks of culture in STEMdiff™-XF RPE Maturation Medium and become fully matured by Day 49, possessing key characteristics such as polygonal shape, polarization, pigmentation and phagocytosis.
Figure 2. Robust and Rapid Generation of Mature Retinal Pigment Epithelial Cells (RPE) across multiple PSC Cell Lines with the STEMdiff™-ACF RPE Differentiation Kit
hPSCs were cultured for 14 Days using STEMdiff™-ACF RPE Differentiation Kit and subsequently subcultured in STEMdiff™-XF RPE Maturation Medium. Flow cytometry expression of RPE markers are shown at Day 14 and Day 49. (A) The percentage of cells expressing PMEL17, RPE65, EZRIN, and CRALBP and (B) Viable cell yields for 4 hPSC cell lines. PMEL17 is expressed at Day 14 and 49 while the other markers are only present at Day 49. Data are reported as mean + SEM; n = 16 -20. (C) A cell pellet of mature RPE cells demonstrates the pigmentation. Maturation is further demonstrated with immunohistochemistry for expression of RPE markers at Day 49. (D, E, F, G) Mature RPE display tight junctions marked by localization of ZO1 and BEST1 to cell junctions. Mature RPE are polarized, expressing EZRIN apically and ZO1 subapically and express proteins required for the visual cycle (RPE65).
Figure 3. Mature Retinal Pigment Epithelial (RPE) Cells Display Key Functionalities Corresponding to RPE Behaviour
hPSC’s were cultured for 14 Days using STEMdiff™-ACF RPE Differentiation Kit and subsequently subcultured on cell culture inserts in STEMdiff™-XF RPE Maturation Medium for 5 weeks. Apical and basal conditioned medium were collected from Mature RPE, and a sandwich ELISA was performed to quantify Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelial Derived Growth Factor (PEDF) secretion. (A, B) Mature RPE secreted more basal VEGF and apical PEDF demonstrating RPE display correct apicobasal polarity. Data shown as mean + SEM; n = 3. (C) Mature RPE were able to generate a strong barrier with high transepithelial resistance (TER). Data shown as mean + SEM; n = 3-6. (D) Mature RPE were fed FITC-labelled bovine photoreceptor outer segments (POS) for 4 to 5 hours prior to being enzymatically dissociated for flow cytometry analysis or fixed with paraformaldehyde for immunostaining. (E) Mature RPE efficiently internalize bovine POS. Data shown as mean + SEM; n = 3. (F) A cross-sectional schematic of the cell insert culture system.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet Product Name
STEMdiff™-XF RPE Maturation Medium Catalog #
100-1365 Lot #
All Language
English Document Type
Safety Data Sheet Product Name
STEMdiff™-XF RPE Maturation Medium Catalog #
100-1365 Lot #
All Language
English Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Research Area
Workflow Stages
Workflow Stages for Human Pluripotent Stem Cell Research
Workflow Stages for hPSC-Derived Sensory Research
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