Collagenase Type IV (1 mg/mL)

Cell dissociation reagent

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Collagenase Type IV (1 mg/mL)

Cell dissociation reagent

100 mL
Catalog #07909
61 USD

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Overview

Human embryonic stem (ES) cells are routinely passaged as clumps, rather than single cells. To generate clumps appropriate for passaging, both enzymatic and mechanical (by dissection or scraping) methods can be used. Collagenase Type IV is an enzyme that is routinely used at 1 mg/mL for the generation of human ES cell and iPS cell clumps for passaging.
Contains:
• 1 mg/mL Collagenase Type IV isolated from Clostridium histolyticum
• DMEM/F-12
Subtype:
Enzymatic
Species:
Human; Mouse; Non-Human Primate; Other; Rat

Technical Resources

Product Documentation

Educational Materials

(2)

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

Publications

(7)
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).
Skeletal Muscle 2015

Dp412e: a novel human embryonic dystrophin isoform induced by BMP4 in early differentiated cells

Massouridès E et al.

Abstract

Background: Duchenne muscular dystrophy (DMD) is a devastating X-linked$nrecessive genetic myopathy. DMD physiopathology is still not fully$nunderstood and a prenatal onset is suspected but difficult to address.$nMethods: The bone morphogenetic protein 4 (BMP4) is a critical signaling$nmolecule involved in mesoderm commitment. Human induced pluripotent stem$ncells (hiPSCs) from DMD and healthy individuals and human embryonic$nstern cells (hESCs) treated with BMP4 allowed us to model the early$nsteps of myogenesis in normal and DMD contexts.$nResults: Unexpectedly, 72h following BMP4 treatment, a new long DMD$ntranscript was detected in all tested hiPSCs and hESCs, at levels$nsimilar to that found in adult skeletal muscle. This novel transcript$nnamed ``Dp412e'' has a specific untranslated first exon which is$nconserved only in a sub group of anthropoids including human. The$ncorresponding novel dystrophin protein of 412-kiloDalton (kDa),$ncharacterized by an N-terminal-truncated actin binding domain, was$ndetected in normal BMP4-treated hiPSCs/hESCs and in embryoid bodies.$nFinally, using a phosphorodiamidate morpholino oligomer (PMO) targeting$nthe DA/ID exon 53, we demonstrated the feasibility of exon skipping$nvalidation with this BMP4-inducible hiPSCs model.$nConclusions: In this study, the use of hiPSCs to analyze early phases of$nhuman development in normal and DMD contexts has led to the discovery of$nan embryonic 412 kDa dystrophin isoform. Deciphering the regulation$nprocess(es) and the function(s) associated to this new isoform can$ncontribute to a better understanding of the DMD physiopathology and$npotential developmental defects. Moreover, the simple and robust$nBMP4-inducible model highlighted here, providing large amount of a long$nDMD transcript and the corresponding protein in only 3 days, is already$nwell adapted to high throughput and high content screening approaches.$nTherefore, availability of this powerful cell platform can accelerate$nthe development, validation and improvement of DMD genetic therapies.
Skeletal muscle 2015

Dp412e: a novel human embryonic dystrophin isoform induced by BMP4 in early differentiated cells.

Massouridès E et al.

Abstract

BACKGROUND Duchenne muscular dystrophy (DMD) is a devastating X-linked recessive genetic myopathy. DMD physiopathology is still not fully understood and a prenatal onset is suspected but difficult to address. METHODS The bone morphogenetic protein 4 (BMP4) is a critical signaling molecule involved in mesoderm commitment. Human induced pluripotent stem cells (hiPSCs) from DMD and healthy individuals and human embryonic stem cells (hESCs) treated with BMP4 allowed us to model the early steps of myogenesis in normal and DMD contexts. RESULTS Unexpectedly, 72h following BMP4 treatment, a new long DMD transcript was detected in all tested hiPSCs and hESCs, at levels similar to that found in adult skeletal muscle. This novel transcript named Dp412e" has a specific untranslated first exon which is conserved only in a sub-group of anthropoids including human. The corresponding novel dystrophin protein of 412-kiloDalton (kDa) characterized by an N-terminal-truncated actin-binding domain was detected in normal BMP4-treated hiPSCs/hESCs and in embryoid bodies. Finally using a phosphorodiamidate morpholino oligomer (PMO) targeting the DMD exon 53 we demonstrated the feasibility of exon skipping validation with this BMP4-inducible hiPSCs model. CONCLUSIONS In this study the use of hiPSCs to analyze early phases of human development in normal and DMD contexts has led to the discovery of an embryonic 412 kDa dystrophin isoform. Deciphering the regulation process(es) and the function(s) associated to this new isoform can contribute to a better understanding of the DMD physiopathology and potential developmental defects. Moreover the simple and robust BMP4-inducible model highlighted here providing large amount of a long DMD transcript and the corresponding protein in only 3 days is already well-adapted to high-throughput and high-content screening approaches. Therefore availability of this powerful cell platform can accelerate the development validation and improvement of DMD genetic therapies."
Nature communications 2014 SEP

Aneuploidy induces profound changes in gene expression, proliferation and tumorigenicity of human pluripotent stem cells.

Ben-David U et al.

Abstract

Human pluripotent stem cells (hPSCs) tend to acquire genomic aberrations in culture, the most common of which is trisomy of chromosome 12. Here we dissect the cellular and molecular implications of this trisomy in hPSCs. Global gene expression analyses reveal that trisomy 12 profoundly affects the gene expression profile of hPSCs, inducing a transcriptional programme similar to that of germ cell tumours. Comparison of proliferation, differentiation and apoptosis between diploid and aneuploid hPSCs shows that trisomy 12 significantly increases the proliferation rate of hPSCs, mainly as a consequence of increased replication. Furthermore, trisomy 12 increases the tumorigenicity of hPSCs in vivo, inducing transcriptionally distinct teratomas from which pluripotent cells can be recovered. Last, a chemical screen of 89 anticancer drugs discovers that trisomy 12 raises the sensitivity of hPSCs to several replication inhibitors. Together, these findings demonstrate the extensive effect of trisomy 12 and highlight its perils for successful hPSC applications.
Cellular reprogramming 2014 OCT

Trisomy 21 mid-trimester amniotic fluid induced pluripotent stem cells maintain genetic signatures during reprogramming: implications for disease modeling and cryobanking.

Pipino C et al.

Abstract

Trisomy 21 is the most common chromosomal abnormality and is associated primarily with cardiovascular, hematological, and neurological complications. A robust patient-derived cellular model is necessary to investigate the pathophysiology of the syndrome because current animal models are limited and access to tissues from affected individuals is ethically challenging. We aimed to derive induced pluripotent stem cells (iPSCs) from trisomy 21 human mid-trimester amniotic fluid stem cells (AFSCs) and describe their hematopoietic and neurological characteristics. Human AFSCs collected from women undergoing prenatal diagnosis were selected for c-KIT(+) and transduced with a Cre-lox-inducible polycistronic lentiviral vector encoding SOX2, OCT4, KLF-4, and c-MYC (50,000 cells at a multiplicity of infection (MOI) 1-5 for 72 h). The embryonic stem cell (ESC)-like properties of the AFSC-derived iPSCs were established in vitro by embryoid body formation and in vivo by teratoma formation in RAG2(-/-), -chain(-/-), C2(-/-) immunodeficient mice. Reprogrammed cells retained their cytogenetic signatures and differentiated into specialized hematopoietic and neural precursors detected by morphological assessment, immunostaining, and RT-PCR. Additionally, the iPSCs expressed all pluripotency markers upon multiple rounds of freeze-thawing. These findings are important in establishing a patient-specific cellular platform of trisomy 21 to study the pathophysiology of the aneuploidy and for future drug discovery.
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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