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TeSR™-E7™重编程培养基(2组分)

无饲养层且不含动物成分的重编程培养基,用于人诱导性多能干细胞(iPS细胞)的诱导。

产品号 #(选择产品)

产品号 #05914_C

无饲养层且不含动物成分的重编程培养基,用于人诱导性多能干细胞(iPS细胞)的诱导。

产品优势

  • 经过预筛选的成分确保iPS细胞克隆形态优良,便于识别并提高人工挑选的效率
  • 减少分化和成纤维细胞的生长,促进均质化iPS细胞培养的快速建立
  • 无饲养层、成分明确的配方有利于高效重复地生成人 iPS 细胞

产品组分包括

  • TeSR™-E7™/ ReproTeSR™ 基础培养基,480 mL
  • TeSR™-E7™ 25X 补充剂,20 mL

总览

TeSR™-E7™(双组分)是一种不含动物成分、成分明确的重编程培养基,经过优化,旨在生成不依赖饲养层的人诱导性多能干细胞(iPS细胞)。该培养基基于Dr. James Thomson(威斯康星大学麦迪逊分校)实验室发布的E7配方。

亚型
专用培养基
 
细胞类型
多能干细胞
 
种属

 
应用
细胞培养,重编程
 
品牌
TeSR
 
研究领域
干细胞生物学
 
制剂类别
Animal Component-Free,无血清,Xeno-Free
 

实验数据

Figure 1. Schematic of Reprogramming Timeline

TeSR™-E7™ can be used during the entire induction phase of reprogramming (day 3 to 25+). Following reprogramming, iPS cell colonies can be isolated and propogated in feeder-free maintenance systems (eg. mTeSR™1 or TeSR™-E8™ media on Corning® Matrigel® or Vitronectin XF™ matrices).

Figure 2. Morphology of Representative iPS Cell Colonies Arising During the Induction Period in TeSR™-E7™

(A-B) Small clusters of colonies with an epithelial-like morphology will appear by one to two weeks following induction (see arrows). (C-D) These clusters expand into pre-iPS cell colonies by two to three weeks. (E-F) Larger ES cell-like colonies are clearly identifiable by three to four weeks. Representative colonies from adult human fibroblasts reprogrammed with episomal vectors containing OCT-4, SOX2, KLF-4, and L-MYC are shown.

Figure 3. Comparison of Primary iPS Cell Colonies Derived Using TeSR™-E7™ and KOSR-Based Medium

(A) TeSR™-E7™ generates colonies with defined borders and less overgrowth of background fibroblasts compared to (B) KOSR-based iPS cell induction medium. Representative colonies from adult human fibroblasts reprogrammed with episomal vectors containing OCT-4, SOX2, KLF-4, and L-MYC are shown.

Figure 4. Comparison of Primary iPS Cell Colonies Derived Using TeSR™-E7™ with Qualified vs Unqualified bFGF

(A) TeSR™-E7™ yields easily recognizable iPS cell colonies with defined borders. (B) Unqualified components can result in colonies that have poorly defined edges and higher levels of differentiation. Representative colonies from adult human fibroblasts reprogrammed with episomal vectors containing OCT-4, SOX2, KLF-4, and L-MYC are shown.

Figure 5. iPS Colonies Expanded in mTeSR™ or TeSR™-E8™

(A - D) iPS cell colonies generated in TeSR™-E7™ and expanded in either mTeSR™1 on Corning® Matrigel® (A-B) or TeSR™-E8™ on Vitronectin XF™ (C, D) exhibit classic ES cell morphology with dense colony centers, defined borders, prominent nucleoli and high nuclear-to-cytoplasmic ratios. (E) iPS cells express high levels of pluripotency markers after just two passages in either mTeSR™1 or TeSR™-E8™ as demonstrated by OCT-4 and SSEA-3 flow cytometry analysis. Data are expressed as mean ± SEM, n = 4.

Figure 6. TeSR™-E7™ Supports Reprogramming of Human Cell Types Including Adult Dermal Fibroblasts and Neonatal Fibroblasts

Reprogramming of (A) adult normal human dermal fibroblasts (NHDF, 33 year-old female) and (B) neonatal foreskin fibroblasts (BJ cells) with episomal reprogramming vectors are shown. TeSR™-E7™ demonstrated similar (in NHDF) or greater (in BJ cells) reprogramming efficiencies compared to KOSR-based iPS cell induction medium. TeSR™-E7™ demonstrated higher reprogramming efficiencies compared to TeSR™-E8™. Data are expressed as mean ± SEM, n ≥ 6, * p ≤ 0.05.

Figure 7. iPS Cells Derived in TeSR™-E7™ Display Normal Karyotype

iPS cell lines were generated in TeSR™-E7™ medium, maintained in mTeSR™1 or TeSR™-E8™ media for a minimum of 5 passages and karyotyped by G-banding karyotype analysis. Three iPS cell lines were analyzed and all demonstrated a normal karyotype; a representative karyogram is shown.

Figure 8. Directed Differentiation of iPS Cells to All Three Germ Layers

TeSR™-E7™-derived iPS cells were differentiated into all three germ layers. Endoderm specification was achieved using the STEMdiff™ Definitive Endoderm Kit, results demonstrated 93.6% SOX17 + CXCR4 + cells. Mesoderm specification was demonstrated using a STEMdiff™ APEL™ medium-based endothelial differentiation protocol, results demonstrated &ht;99% CD31 + cells (data not shown) and 84.8% VEGFR2 + CD105 + cells. Ectoderm specification was demonstrated using STEMdiff™ Neural Induction Medium, immunocytochemistry shows high levels of PAX6 staining with no detectable OCT-4 staining by day 9 of neural induction.

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明,或浏览下方更多实验方案。

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
05914
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
05914
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
05914
Lot #
All
Language
English

应用领域

本产品专为以下研究领域设计,适用于工作流程中的高亮阶段。探索这些工作流程,了解更多我们为各研究领域提供的其他配套产品。

相关材料与文献

技术资料 (10)

文献 (1)

Disruption of GRIN2B Impairs Differentiation in Human Neurons. S. Bell et al. Stem cell reports 2018 JUL

Abstract

Heterozygous loss-of-function mutations in GRIN2B, a subunit of the NMDA receptor, cause intellectual disability and language impairment. We developed clonal models of GRIN2B deletion and loss-of-function mutations in a region coding for the glutamate binding domain in human cells and generated neurons from a patient harboring a missense mutation in the same domain. Transcriptome analysis revealed extensive increases in genes associated with cell proliferation and decreases in genes associated with neuron differentiation, a result supported by extensive protein analyses. Using electrophysiology and calcium imaging, we demonstrate that NMDA receptors are present on neural progenitor cells and that human mutations in GRIN2B can impair calcium influx and membrane depolarization even in a presumed undifferentiated cell state, highlighting an important role for non-synaptic NMDA receptors. It may be this function, in part, which underlies the neurological disease observed in patients with GRIN2B mutations.

更多信息

更多信息
种属 Human
配方类别 Animal Component-Free, Serum-Free, Xeno-Free
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