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STEMdiff™-ACF RPE 分化试剂盒

无动物成分的分化试剂盒，用于从人多能干细胞生成未成熟的视网膜色素上皮细胞
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产品号 #100-1367_C

无动物成分的分化试剂盒，用于从人多能干细胞生成未成熟的视网膜色素上皮细胞

产品优势

  • 采用无动物成分和无血清配方，确保结果一致性
  • 简化 RPE 分化流程，具有简单、可扩展的工作流程
  • 在仅需 14 天内即可从 hPSC 生成未成熟的 RPE 细胞
  • 可分化出高纯度、功能性 RPE 细胞，已在多种人类胚胎干细胞和诱导多能干细胞系中验证
  • 提供便捷、用户友好且可扩展的操作流程，增强实验信心
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Need a high-quality cell source? Use the hiPSC SCTi003-A (female) or SCTi004-A (male) control lines, manufactured with mTeSR™ Plus.
总览
实验数据
产品说明书及文档
应用领域
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总览

使用 STEMdiff™-ACF RPE 分化试剂盒，可在 14 天内快速且高效地生成 hPSC 衍生的未成熟视网膜色素上皮 (RPE)。该试剂盒不含动物成分和血清，可在mTeSR™1或mTeSR™ Plus中培养 hPSC 细胞，生成超过 50% PMEL17 阳性的未成熟 RPE 细胞。

使用 STEMdiff™-ACF RPE 分化试剂盒生成的未成熟 RPE 可以作为中间细胞库冻存，或使用STEMdiff™-XF RPE 成熟培养基进一步成熟为功能性 RPE 。通过将这些产品结合使用，无需手动筛选或细胞富集，最快 49 天即可从 hPSC 生成高纯度 RPE 细胞群（图 2）。在此工作流程中添加STEMdiff™-ACF RPE 接种添加剂，可提高 RPE 细胞分化成熟的产量和存活率。 使用这些产品获得的细胞可用于模拟人类视网膜发育和疾病、药物筛选、细胞和基因治疗验证以及高级组织模型开发。

如果您计划将此产品用于商业或临床应用，请联系我们。

细胞类型
神经细胞，PSC衍生，多能干细胞
 
应用
细胞培养，分化
 
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
 

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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 RPE Kit

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).

Mature Retinal Pigment Epithelial (RPE) Cells Display Key Functionalities Corresponding to RPE Behaviour

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™-ACF RPE Differentiation Kit
Catalog #
100-1367
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
STEMdiff™-ACF RPE Differentiation Kit
Catalog #
100-1367
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
STEMdiff™-ACF RPE Differentiation Kit
Catalog #
100-1367
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
STEMdiff™-ACF RPE Differentiation Kit
Catalog #
100-1367
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
STEMdiff™-ACF RPE Differentiation Kit
Catalog #
100-1367
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
STEMdiff™-ACF RPE Differentiation Kit
Catalog #
100-1367
Lot #
All
Language
English
Document Type
Safety Data Sheet 6
Product Name
STEMdiff™-ACF RPE Differentiation Kit
Catalog #
100-1367
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
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
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ISO 13485 和 ISO 9001 质量管理体系认证

ISO 14001 环境管理体系认证

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