TeSR™-E7™

Feeder-free, xeno-free reprogramming medium for human iPS cell induction

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TeSR™-E7™ Medium for Reprogramming

Feeder-free, xeno-free reprogramming medium for human iPS cell induction

500 mL
Catalog #05910
221 USD

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Overview

TeSR™-E7™ is a xeno-free and defined reprogramming culture medium optimized for the generation of human iPS cells without the use of feeders. It is based on the E7 formulation published by the laboratory of Dr. James Thomson (University of Wisconsin-Madison).
Advantages:
• Pre-screened components ensure high quality iPS cell colony morphology for easy identification and improved manual selection
• Reduced differentiation and fibroblast growth enables rapid establishment of homogeneous iPS cell cultures
• Feeder-free, defined formulation facilitates reproducibly efficient human iPS cell generation
Components:
  • TeSR™-E7™ Basal Medium, 474 mL
  • TeSR™-E7™ 20X Supplement, 25 mL
  • TeSR™-E7™ 500X Supplement, 1 mL
Subtype:
Specialized Media
Cell Type:
Pluripotent Stem Cells
Species:
Human
Application:
Reprogramming; Cell Culture
Brand:
TeSR
Area of Interest:
Stem Cell Biology
Formulation:
Serum-Free; Xeno-Free; Defined

Technical Resources

Educational Materials

(6)

Product 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.

Data and Publications

Data

Schematic of Reprogramming Timeline

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).

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

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.

Comparison of Primary iPS Cell Colonies Derived Using TeSR™-E7™ and KOSR-based Medium

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.

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

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.

iPS Colonies Expanded in mTeSR™ or TeSR™-E8™

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.

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

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.

iPS Cells Derived in TeSR™-E7™ Display Normal Karyotype

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.

Directed Differentiation of iPS Cells to All Three Germ Layers

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.

Publications

(6)
Scientific reports 2016 MAR

TGF signaling regulates the choice between pluripotent and neural fates during reprogramming of human urine derived cells.

Wang L et al.

Abstract

Human urine cells (HUCs) can be reprogrammed into neural progenitor cells (NPCs) or induced pluripotent stem cells (iPSCs) with defined factors and a small molecule cocktail, but the underlying fate choice remains unresolved. Here, through sequential removal of individual compound from small molecule cocktail, we showed that A8301, a TGF signaling inhibitor, is sufficient to switch the cell fate from iPSCs into NPCs in OSKM-mediated HUCs reprogramming. However, TGF exposure at early stage inhibits HUCs reprogramming by promoting EMT. Base on these data, we developed an optimized approach for generation of NPCs or iPSCs from HUCs with significantly improved efficiency by regulating TGF activity at different reprogramming stages. This approach provides a simplified and improved way for HUCs reprogramming, thus would be valuable for banking human iPSCs or NPCs from people with different genetic background.
Oncology reports 2016 AUG

Regulation of tumorigenesis in oral epithelial cells by defined reprogramming factors Oct4 and Sox2.

Cai J et al.

Abstract

Oct4 and Sox2 are pluripotent stem cell factors but the interplay between them in tumorigenesis is unclear. The aim of the present study was to investigate the roles of Oct4 and Sox2 in the reprogramming of oral cancer stem cells. One or both Oct4 and Sox2 were overexpressed in immortalized oral epithelial (hTERT+-OME) cells by lentivirus transduction. In addition, Oct4 and Sox2 proteins in two oral squamous cell carcinoma cell (OSCC) lines (Cal27 and primary cultured OSCC from a T2N2M0 patient) were individually or combinedly knocked down by shRNA. The results showed that the doubly transduced (Oct4+Sox2+) cells could trigger neoplasms in immunodeficient mice after lentivirus transduction, but single transduced (Oct4+ or Sox2+) cells had no tumor formation ability. The knockdown Sox2low and knockdown Oct4lowSox2low cells resulted in decreased tumor size in the immunodeficient mice but the single knockdown Oct4low cancer cells acquired more aggressive xenografts. Our findings suggest that Oct4+Sox2+ cells may be reprogrammed cancer stem cells inducing oral carcinogenesis.
Stem Cells International 2016

Utility of Lymphoblastoid Cell Lines for Induced Pluripotent Stem Cell Generation

Kumar S et al.

Abstract

A large number of EBV immortalized LCLs have been generated and maintained in genetic/epidemiological studies as a perpetual source of DNA and as a surrogate in vitro cell model. Recent successes in reprograming LCLs into iPSCs have paved the way for generating more relevant in vitro disease models using this existing bioresource. However, the overall reprogramming efficiency and success rate remain poor and very little is known about the mechanistic changes that take place at the transcriptome and cellular functional level during LCL-to-iPSC reprogramming. Here, we report a new optimized LCL-to-iPSC reprogramming protocol using episomal plasmids encoding pluripotency transcription factors and mouse p53DD (p53 carboxy-terminal dominant-negative fragment) and commercially available reprogramming media. We achieved a consistently high reprogramming efficiency and 100% success rate using this optimized protocol. Further, we investigated the transcriptional changes in mRNA and miRNA levels, using FC-abs ≥ 2.0 and FDR ≤ 0.05 cutoffs; 5,228 mRNAs and 77 miRNAs were differentially expressed during LCL-to-iPSC reprogramming. The functional enrichment analysis of the upregulated genes and activation of human pluripotency pathways in the reprogrammed iPSCs showed that the generated iPSCs possess transcriptional and functional profiles very similar to those of human ESCs.
PLoS ONE 2016

Multiple sclerosis patient-specific primary neurons differentiated from urinary renal epithelial cells via induced pluripotent stem cells

Massa MG et al.

Abstract

As multiple sclerosis research progresses, it is pertinent to continue to develop suitable paradigms to allow for ever more sophisticated investigations. Animal models of multiple sclerosis, despite their continuing contributions to the field, may not be the most prudent for every experiment. Indeed, such may be either insufficient to reflect the functional impact of human genetic variations or unsuitable for drug screenings. Thus, we have established a cell- and patient-specific paradigm to provide an in vitro model within which to perform future genetic investigations. Renal proximal tubule epithelial cells were isolated from multiple sclerosis patients' urine and transfected with pluripotency-inducing episomal factors. Subsequent induced pluripotent stem cells were formed into embryoid bodies selective for ectodermal lineage, resulting in neural tube-like rosettes and eventually neural progenitor cells. Differentiation of these precursors into primary neurons was achieved through a regimen of neurotrophic and other factors. These patient-specific primary neurons displayed typical morphology and functionality, also staining positive for mature neuronal markers. The development of such a non-invasive procedure devoid of permanent genetic manipulation during the course of differentiation, in the context of multiple sclerosis, provides an avenue for studies with a greater cell- and human-specific focus, specifically in the context of genetic contributions to neurodegeneration and drug discovery.
Nature protocols 2012 NOV

Passaging and colony expansion of human pluripotent stem cells by enzyme-free dissociation in chemically defined culture conditions.

Beers J et al.

Abstract

This protocol describes an EDTA-based passaging procedure to be used with chemically defined E8 medium that serves as a tool for basic and translational research into human pluripotent stem cells (PSCs). In this protocol, passaging one six-well or 10-cm plate of cells takes about 6-7 min. This enzyme-free protocol achieves maximum cell survival without enzyme neutralization, centrifugation or drug treatment. It also allows for higher throughput, requires minimal material and limits contamination. Here we describe how to produce a consistent E8 medium for routine maintenance and reprogramming and how to incorporate the EDTA-based passaging procedure into human induced PSC (iPSC) derivation, colony expansion, cryopreservation and teratoma formation. This protocol has been successful in routine cell expansion, and efficient for expanding large-volume cultures or a large number of cells with preferential dissociation of PSCs. Effective for all culture stages, this procedure provides a consistent and universal approach to passaging human PSCs in E8 medium.
STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.
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