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: 41 USD

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

Scientific 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

(10)
Cell 2018 JAN

Intrinsic Immunity Shapes Viral Resistance of Stem Cells.

Wu X et al.

Abstract

Stem cells are highly resistant to viral infection compared to their differentiated progeny; however, the mechanism is mysterious. Here, we analyzed gene expression in mammalian stem cells and cells at various stages of differentiation. We find that, conserved across species, stem cells express a subset of genes previously classified as interferon (IFN) stimulated genes (ISGs) but that expression is intrinsic, as stem cells are refractory to interferon. This intrinsic ISG expression varies in a cell-type-specific manner, and many ISGs decrease upon differentiation, at which time cells become IFN responsive, allowing induction of a broad spectrum of ISGs by IFN signaling. Importantly, we show that intrinsically expressed ISGs protect stem cells against viral infection. We demonstrate the in vivo importance of intrinsic ISG expression for protecting stem cells and their differentiation potential during viral infection. These findings have intriguing implications for understanding stem cell biology and the evolution of pathogen resistance.
Cell stem cell 2017 APR

Human iPSC-Derived Cerebral Organoids Model Cellular Features of Lissencephaly and Reveal Prolonged Mitosis of Outer Radial Glia.

Bershteyn M et al.

Abstract

Classical lissencephaly is a genetic neurological disorder associated with mental retardation and intractable epilepsy, and Miller-Dieker syndrome (MDS) is the most severe form of the disease. In this study, to investigate the effects of MDS on human progenitor subtypes that control neuronal output and influence brain topology, we analyzed cerebral organoids derived from control and MDS-induced pluripotent stem cells (iPSCs) using time-lapse imaging, immunostaining, and single-cell RNA sequencing. We saw a cell migration defect that was rescued when we corrected the MDS causative chromosomal deletion and severe apoptosis of the founder neuroepithelial stem cells, accompanied by increased horizontal cell divisions. We also identified a mitotic defect in outer radial glia, a progenitor subtype that is largely absent from lissencephalic rodents but critical for human neocortical expansion. Our study, therefore, deepens our understanding of MDS cellular pathogenesis and highlights the broad utility of cerebral organoids for modeling human neurodevelopmental disorders.
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.
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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