ReLeSR™

Enzyme-free human ES and iPS cell selection and passaging reagent

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Enzyme-free human ES and iPS cell selection and passaging reagent
From: 40 USD

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Overview

ReLeSR™ is an enzyme-free reagent for dissociation and passaging of human pluripotent stem cells (hPSCs) as aggregates without manual selection or scraping. Passaging hPSCs with ReLeSR™ easily generates optimally-sized aggregates, while eliminating the hassle and variability associated with manual manipulation. By eliminating the need for scraping, ReLeSR™ enables the use of culture flasks and other closed vessels, thus facilitating culture scale-up and automation.
Advantages:
• Simple passaging protocol


• Eliminates the need for manual removal (selection) of differentiated cells


• No manual scraping to generate cell aggregates
• Compatible with passaging in flasks and large culture vessels
• Chemically defined, enzyme-free, and gentle on cells
• High expansion of human ES/iPS cells after passaging
Subtype:
Non-Enzymatic
Cell Type:
Pluripotent Stem Cells
Species:
Human
Brand:
ReLeSR; TeSR
Area of Interest:
Stem Cell Biology

Technical Resources

Product Documentation

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Educational Materials

(7)

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

Passaging Protocol Comparison

Figure 1. Passaging Protocol Comparison

ReLeSR™ passaging protocol eliminates difficult and time-consuming steps, thereby enabling easy culture scale-up.
Surface area of 4 x 6 well plates (230 cm²) is comparable to that of a T225 flask (225 cm²).
TeSR™ = TeSR™ family media (mTeSR™1, TeSR™2, or TeSR™-E8™).

Selectively Detach Undifferentiated Cells

Figure 2. Selectively Detach Undifferentiated Cells

ReLeSR™ selectively detaches undifferentiated cells from pluripotent stem cell cultures without manual selection. Optimally-sized aggregates are generated following shaking/tapping of the cultureware.
(A) An hPSC culture ready for passaging. Note the presence of differentiated cells at the edge of the undifferentiated hPSC colony. (B) Following incubation with ReLeSR™, the undifferentiated hPSC colony starts to lift off of the cultureware. The differentiated cells remain attached to the cultureware. (C) Following shaking/tapping of the cultureware, the undifferentiated cells completely lift off of the cultureware. (D) The undifferentiated hPSC colony is broken up into optimally-sized aggregates for replating.

Rescue Highly Differentiated Cultures

Figure 3. Rescue Highly Differentiated Cultures

Poor quality human pluripotent stem cell cultures containing large proportions of differentiated cells can be rescued by passaging with ReLeSR™. (A) A poor quality hPSC culture containing ~50% undifferentiated cells. (B) Following passaging with ReLeSR™, the differentiated cells have largely been eliminated from the culture, with >90% undifferentiated cells present at the end of the next passage.

Select Putative iPS Cell Clones

Figure 4. Select Putative iPS Cell Clones

Easily isolate newly generated human iPS cell colonies with ReLeSR™ by selectively detaching undifferentiated cells and leaving non reprogrammed cells behind.
(A) A TeSR™-E7™ reprogramming culture which has been treated with ReLeSR™ to detach the putative iPS cell colony, leaving the non-reprogrammed and differentiated cells behind. (B) Cultures contain a high proportion of undifferentiated cells by the end of the first passage.

Publications

(6)
EMBO molecular medicine 2016 OCT

Coenzyme A corrects pathological defects in human neurons of PANK2-associated neurodegeneration.

Orellana DI et al.

Abstract

Pantothenate kinase-associated neurodegeneration (PKAN) is an early onset and severely disabling neurodegenerative disease for which no therapy is available. PKAN is caused by mutations in PANK2, which encodes for the mitochondrial enzyme pantothenate kinase 2. Its function is to catalyze the first limiting step of Coenzyme A (CoA) biosynthesis. We generated induced pluripotent stem cells from PKAN patients and showed that their derived neurons exhibited premature death, increased ROS production, mitochondrial dysfunctions-including impairment of mitochondrial iron-dependent biosynthesis-and major membrane excitability defects. CoA supplementation prevented neuronal death and ROS formation by restoring mitochondrial and neuronal functionality. Our findings provide direct evidence that PANK2 malfunctioning is responsible for abnormal phenotypes in human neuronal cells and indicate CoA treatment as a possible therapeutic intervention.
Cell reports 2016 NOV

Nucleosome Density ChIP-Seq Identifies Distinct Chromatin Modification Signatures Associated with MNase Accessibility.

Lorzadeh A et al.

Abstract

Nucleosome position, density, and post-translational modification are widely accepted components of mechanisms regulating DNA transcription but still incompletely understood. We present a modified native ChIP-seq method combined with an analytical framework that allows MNase accessibility to be integrated with histone modification profiles. Application of this methodology to the primitive (CD34+) subset of normal human cord blood cells enabled genomic regions enriched in one versus two nucleosomes marked by histone 3 lysine 4 trimethylation (H3K4me3) and/or histone 3 lysine 27 trimethylation (H3K27me3) to be associated with their transcriptional and DNA methylation states. From this analysis, we defined four classes of promoter-specific profiles and demonstrated that a majority of bivalent marked promoters are heterogeneously marked at a single-cell level in this primitive cell type. Interestingly, extension of this approach to human embryonic stem cells revealed an altered relationship between chromatin modification state and nucleosome content at promoters, suggesting developmental stage-specific organization of histone methylation states.
Stem Cell Reports 2016 MAY

Intercellular Adhesion-Dependent Cell Survival and ROCK-Regulated Actomyosin-Driven Forces Mediate Self-Formation of a Retinal Organoid

Lowe A et al.

Abstract

In this study we dissected retinal organoid morphogenesis in human embryonic stem cell (hESC)-derived cultures and established a convenient method for isolating large quantities of retinal organoids for modeling human retinal development and disease. Epithelialized cysts were generated via floating culture of clumps of Matrigel/hESCs. Upon spontaneous attachment and spreading of the cysts, patterned retinal monolayers with tight junctions formed. Dispase-mediated detachment of the monolayers and subsequent floating culture led to self-formation of retinal organoids comprising patterned neuroretina, ciliary margin, and retinal pigment epithelium. Intercellular adhesion-dependent cell survival and ROCK-regulated actomyosin-driven forces are required for the self-organization. Our data supports a hypothesis that newly specified neuroretina progenitors form characteristic structures in equilibrium through minimization of cell surface tension. In long-term culture, the retinal organoids autonomously generated stratified retinal tissues, including photoreceptors with ultrastructure of outer segments. Our system requires minimal manual manipulation, has been validated in two lines of human pluripotent stem cells, and provides insight into optic cup invagination in vivo.
Nature Materials 2016 MAR

Defined three-dimensional microenvironments boost induction of pluripotency

Caiazzo M et al.

Abstract

Since the discovery of induced pluripotent stem cells (iPSCs), numerous approaches have been explored to improve the original protocol, which is based on a two-dimensional (2D) cell-culture system. Surprisingly, nothing is known about the effect of a more biologically faithful 3D environment on somatic-cell reprogramming. Here, we report a systematic analysis of how reprogramming of somatic cells occurs within engineered 3D extracellular matrices. By modulating microenvironmental stiffness, degradability and biochemical composition, we have identified a previously unknown role for biophysical effectors in the promotion of iPSC generation. We find that the physical cell confinement imposed by the 3D microenvironment boosts reprogramming through an accelerated mesenchymal-to-epithelial transition and increased epigenetic remodelling. We conclude that 3D microenvironmental signals act synergistically with reprogramming transcription factors to increase somatic plasticity.
Methods in molecular biology (Clifton, N.J.) 2016 FEB

A Concise Protocol for siRNA-Mediated Gene Suppression in Human Embryonic Stem Cells.

Renz PF and Beyer TA

Abstract

Human embryonic stem cells hold great promise for future biomedical applications such as disease modeling and regenerative medicine. However, these cells are notoriously difficult to culture and are refractory to common means of genetic manipulation, thereby limiting their range of applications. In this protocol, we present an easy and robust method of gene repression in human embryonic stem cells using lipofection of small interfering RNA (siRNA).
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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