STEMdiff™ Trilineage Differentiation Kit

Functional assay kit to assess pluripotency by directed differentiation of human ES and iPS cells to all three germ layers
STEMdiff™ Trilineage Differentiation Kit

Functional assay kit to assess pluripotency by directed differentiation of human ES and iPS cells to all three germ layers

1 Kit
Catalog # 05230
Required Products
  1. Gentle Cell Dissociation Reagent
    Gentle Cell Dissociation Reagent

    cGMP, enzyme-free cell dissociation reagent

  2. ACCUTASE™
    ACCUTASE™

    Cell detachment solution

  3. Y-27632 (Dihydrochloride)
    Y-27632

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

  4. mTeSR™1
    mTeSR™1

    cGMP, feeder-free maintenance medium for human ES and iPS cells

Overview

The STEMdiff™ Trilineage Differentiation Kit provides a simple culture assay to functionally validate the ability of new or established human embryonic stem (ES) and induced pluripotent stem (iPS) cell lines to differentiate to the three germ layers: ectoderm, mesoderm, and endoderm. This kit includes specialized, complete media and monolayer-based protocols to perform parallel in vitro directed differentiation experiments for each germ layer, clearly and reproducibly establishing trilineage differentiation potential within one week. STEMdiff™ Trilineage Differentiation Kit is intended to be an endpoint assay and is not optimized for the generation of cells for downstream differentiation or other applications. STEMdiff™ Trilineage Differentiation Kit has been optimized to assess cells maintained in mTeSR™1.
Advantages
• Reproducible differentiation to all three germ layers across multiple pluripotent cell lines
• Easy-to-interpret assay results
• Complete, defined culture media
• Standardized, one-week protocol
Components
  • STEMdiff™ Trilineage Ectoderm Medium, 175 mL
  • STEMdiff™ Trilineage Mesoderm Medium, 100 mL
  • STEMdiff™ Trilineage Endoderm Medium, 100 mL
Subtype
Specialized Media
Cell Type
Pluripotent Stem Cells
Species
Human
Application
Cell Culture, Characterization, Differentiation, Functional Assay, Phenotyping
Brand
STEMdiff
Area of Interest
Stem Cell Biology

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
STEMdiff™ Trilineage Differentiation Kit
Catalog #
05230
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
STEMdiff™ Trilineage Differentiation Kit
Catalog #
05230
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
STEMdiff™ Trilineage Differentiation Kit
Catalog #
05230
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
STEMdiff™ Trilineage Differentiation Kit
Catalog #
05230
Lot #
All
Language
English

Educational Materials (27)

Brochure
STEMdiff™ Trilineage Differentiation Kit
Brochure
qPCR Arrays for Cell Characterization
Brochure
2019-2020 Cell Culture Training Catalog
Brochure
Products for Human Pluripotent Stem Cells
Wallchart
Directed Differentiation of Pluripotent Stem Cells
Wallchart
Derivation and Applications of Human Pluripotent Stem Cells
Video
How to Coat Plates for Human Pluripotent Stem Cell (hPSC) Cultures in mTeSR™ Plus
5:20
How to Coat Plates for Human Pluripotent Stem Cell (hPSC) Cultures in mTeSR™ Plus
Video
How to Generate Cell Aggregates and Passage Human Pluripotent Stem Cells (hPSCs) in mTeSR™ Plus
7:57
How to Generate Cell Aggregates and Passage Human Pluripotent Stem Cells (hPSCs) in mTeSR™ Plus
Video
How to Set Up an Assay with the hPSC Genetic Analysis Kit Experiment
8:33
How to Set Up an Assay with the hPSC Genetic Analysis Kit Experiment
Video
How to Maintain and Assess Morphology of Human Pluripotent Stem Cells (hPSCs) in mTeSR™ Plus
3:35
How to Maintain and Assess Morphology of Human Pluripotent Stem Cells (hPSCs) in mTeSR™ Plus
Video
How to Transition Human Pluripotent Stem Cells into mTeSR™ Plus from Other Feeder-Free Media
1:29
How to Transition Human Pluripotent Stem Cells into mTeSR™ Plus from Other Feeder-Free Media
Webinar
Nature Research Round Table: Maintenance of Human Pluripotent Stem Cells In Vitro
20:09
Nature Research Round Table: Maintenance of Human Pluripotent Stem Cells In Vitro
Webinar
hPSC Quality: Essential Considerations for Gene Editing, Cloning, Maintenance and Disease Modeling
50:49
hPSC Quality: Essential Considerations for Gene Editing, Cloning, Maintenance and Disease Modeling
Webinar
STEMdiff™ Kits for Robust and Efficient Differentiation of hPSCs to Multiple Cell Types
20:24
STEMdiff™ Kits for Robust and Efficient Differentiation of hPSCs to Multiple Cell Types
Webinar
Survey Results: Insights and Trends in Pluripotent Stem Cell Research
13:58
Survey Results: Insights and Trends in Pluripotent Stem Cell Research
Webinar
Considerations for High-Efficiency Genome Editing of Human Pluripotent Stem Cells
50:01
Considerations for High-Efficiency Genome Editing of Human Pluripotent Stem Cells
Webinar
Nature Research Round Table: Defining and Maintaining Pluripotency & hPSC Line Registration and Banking - Panel Discussion
33:47
Nature Research Round Table: Defining and Maintaining Pluripotency & hPSC Line Registration and Banking - Panel Discussion
Webinar
Development, Compatibility, and Applications of mTeSR™ Plus; an Enhanced Medium for the Maintenance of Human Pluripotent Stem Cells (hPSCs)
42:10
Development, Compatibility, and Applications of mTeSR™ Plus; an Enhanced Medium for the Maintenance of Human Pluripotent Stem Cells (hPSCs)
Webinar
Using CRISPR/Cas9 to Model Stem Cell Organization and Dynamics
71:57
Using CRISPR/Cas9 to Model Stem Cell Organization and Dynamics
Webinar
iPSCs As Models, Part 1: What Are Induced Pluripotent Stem Cells and How Do You Use Them?
37:18
iPSCs As Models, Part 1: What Are Induced Pluripotent Stem Cells and How Do You Use Them?
Webinar
Quality Control Guidelines for Clinical-Grade Human Induced Pluripotent Stem Cell Lines
1:13:44
Quality Control Guidelines for Clinical-Grade Human Induced Pluripotent Stem Cell Lines
Webinar
Best Practice Criteria for Pluripotent Stem Cell Lines
55:52
Best Practice Criteria for Pluripotent Stem Cell Lines
Webinar
Nature Research Round Table: Pluripotency Tests
17:46
Nature Research Round Table: Pluripotency Tests
Webinar
Maintaining and Assessing High-Quality hPSC Cultures
55:53
Maintaining and Assessing High-Quality hPSC Cultures
Webinar
Human Pluripotent Stem Cells for the Treatment of Age-Related Macular Degeneration and Compliance Considerations for Clinical-Grade iPSCs
47:23
Human Pluripotent Stem Cells for the Treatment of Age-Related Macular Degeneration and Compliance Considerations for Clinical-Grade iPSCs
Webinar
Improving Reproducibility of Your hPSC Research by Generating a High-Quality Cell Bank
21:53
Improving Reproducibility of Your hPSC Research by Generating a High-Quality Cell Bank
Scientific Poster
Rapid Assessment of Pluripotency Using Directed Differentiation to all Three Germ Layers
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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 (3)

Stem cells and development 2017 MAY Feeder-Free Derivation of Naïve Human Pluripotent Stem Cells. Ward E et al.

Abstract

Human pluripotent stem cells (HPSCs) cultured in conditions that maintain pluripotency via FGF and TGFβ signaling have been described as being in a primed state. These cells have been shown to exhibit characteristics more closely related to mouse epiblast-derived stem cells than to so called naïve mouse PSCs said to possess a more ground state pluripotency that mimics the early mouse embryo inner cell mass. Initial attempts to create culture conditions favorable for generation of naïve HPSCs from primed HPSCs has required the use of mouse embryonic fibroblasts as a feeder layer to support this transition. A protocol for the routine derivation and maintenance of naïve HPSCs in completely defined conditions is highly desirable for stem cell researchers to enhance the study and clinical translation of naïve HPSCs. Here we describe a standard protocol for transitioning primed HPSCs to a naïve state using commercial RSet media and xeno-free recombinant vitronectin.
Stem cell research 2017 Peripheral blood derived induced pluripotent stem cells (iPSCs) from a female with familial hypertrophic cardiomyopathy. S. B. Ross et al.

Abstract

Induced pluripotent stem cells (iPSCs) were generated from peripheral blood mononuclear cells (PBMCs) obtained from a 62-year-old female with familial hypertrophic cardiomyopathy (HCM). PBMCs were reprogrammed to a pluripotent state following transfection with non-integrative episomal vectors carrying reprogramming factors OCT4, SOX2, LIN28, KLF4 and L-MYC. iPSCs were shown to express pluripotency markers, possess trilineage differentiation potential, carry rare variants identified in DNA isolated directly from the patient's whole blood, have a normal karyotype and no longer carry episomal vectors for reprogramming. This line is a useful resource for identifying unknown genetic causes of HCM.
Stem cell research 2017 Generation of induced pluripotent stem cells (iPSCs) from a hypertrophic cardiomyopathy patient with the pathogenic variant p.Val698Ala in beta-myosin heavy chain (MYH7) gene. S. B. Ross et al.

Abstract

Induced pluripotent stem cells (iPSCs) were generated from peripheral blood mononuclear cells (PBMCs) isolated from the whole blood of a 43-year-old male with hypertrophic cardiomyopathy (HCM) who carries the pathogenic variant p.Val698Ala in beta-myosin heavy chain (MYH7). Patient-derived PBMCs were reprogrammed using non-integrative episomal vectors containing reprogramming factors OCT4, SOX2, LIN28, KLF4 and L-MYC. iPSCs were shown to express pluripotent markers, have trilineage differentiation potential, carry the pathogenic MYH7 variant p.Val698Ala, have a normal karyotype and no longer carry the episomal reprogramming vector. This line is useful for studying the link between variants in MYH7 and the pathogenesis of HCM.
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