STEMdiff™ Trilineage Differentiation Assay Kit

STEMdiff™ Trilineage Differentiation Kit

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

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STEMdiff™ Trilineage Differentiation Kit

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

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

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Product 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

What's Included

STEMdiff™ Trilineage Ectoderm Medium, 175 mL

STEMdiff™ Trilineage Mesoderm Medium, 100 mL

STEMdiff™ Trilineage Endoderm Medium, 100 mL

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To see all required products for your protocol, please consult the Protocols and Documentation.

Gentle Cell Dissociation Reagent
cGMP, enzyme-free cell dissociation reagent
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Cell detachment solution
Y-27632 (Dihydrochloride)
RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2
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cGMP, feeder-free maintenance medium for human ES and iPS cells
Labeling Antibodies
Compatible antibodies for purity assessment of isolated 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 ��ձ𳧸�™1, mTeSR™ Plus, or TeSR™- AOF.

Specialized Media

Pluripotent Stem Cells

Human

Cell Culture, Characterization, Differentiation, Functional Assay, Phenotyping

STEMdiff

Stem Cell Biology

More Information

More Information
Safety Statement	CA WARNING: This product can expose you to chemicals including Nickel Compounds which are known to the State of California to cause cancer and birth defects or other reproductive harm. For more information go to

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type

Product Name

Catalog #

Lot #

Language

Document Type

Product Name

Catalog #

05230

Lot #

All

Language

English

Document Type

Product Name

Catalog #

05230

Lot #

All

Language

English

Document Type

Product Name

Catalog #

05230

Lot #

All

Language

English

Document Type

Product Name

Catalog #

05230

Lot #

All

Language

English

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.

Research Area

Workflow Stages

Workflow Stages for Human Pluripotent Stem Cell Research

Reprogramming

Maintenance

Differentiation

Workflow Stages for PSC Quality Control

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Workflow Stages for hPSC-Derived Cell Therapy Development

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Resources and Publications

Functional characterization of the MED12 p.Arg1138Trp variant in females: implications for neural development and disease mechanism N. C. Shaw et al. Molecular Medicine 2025 Sep

Abstract

Seven female individuals with multiple congenital anomalies, developmental delay and/or intellectual disability have been found to have a genetic variant of uncertain significance in the mediator complex subunit 12 gene ( MED12 c.3412C>T, p.Arg1138Trp). The functional consequence of this genetic variant in disease is undetermined, and insight into disease mechanism is required. We identified a de novo MED12 p.Arg1138Trp variant in a female patient and compared disease phenotypes with six female individuals identified in the literature. To investigate affected biological pathways, we derived two induced pluripotent stem cell (iPSC) lines from the patient: one expressing wildtype MED12 and the other expressing the MED12 p.Arg1138Trp variant. We performed neural disease modelling, transcriptomics and protein analysis, comparing healthy and variant cells. When comparing the two cell lines, we identified altered gene expression in neural cells expressing the variant, including genes regulating RNA polymerase II activity, transcription, pre-mRNA processing, and neural development. We also noted a decrease in MED12L expression. Pathway analysis indicated temporal delays in axon development, forebrain differentiation, and neural cell specification with significant upregulation of pre-ribosome complex gene pathways. In a human neural model, expression of MED12 p.Arg1138Trp altered neural cell development and dysregulated the pre-ribosome complex providing functional evidence of disease aetiology and mechanism in MED12-related disorders. The online version contains supplementary material available at 10.1186/s10020-025-01365-5.

Generation of human induced pluripotent stem cell lines from patients with FGFR2 -linked syndromic craniosynostosis M. Gijsbertsen et al. Disease Models & Mechanisms 2025 Sep

Abstract

Craniosynostosis is a multigenic congenital condition in which one or more calvarial sutures have prematurely fused during the development of the fetus. Pathogenic variants in FGFR2 are associated with the development of syndromic craniosynostosis, such as Crouzon, Apert and Pfeifer syndromes. Investigation of FGFR2 -linked craniosynostosis is hindered by the lack of appropriate in vitro models. Patient-derived human induced pluripotent stem cell (hiPSC) in vitro disease models provide the opportunity to investigate the disease, identify molecular targets for pharmaceutical treatments, and enable the generation of autologous pluripotent stem cell catalogues. Here, we report three patient-derived hiPSC lines carrying the C342Y, S252W or E565G FGFR2 pathogenic variant. The patient hiPSC lines express characteristic pluripotency markers and display distinct phosphorylation profiles under unstimulated conditions. FGFR2 C342Y showed autophosphorylation in the absence of bFGF ligand, although downstream docking proteins PLCγ and FRS2α were not phosphorylated. FGFR2 S252W and FGFR2 E565G hiPSCs showed increased phosphorylation of docking proteins PLCγ and FRS2α, whereas FGFR2 was not phosphorylated. These patient hiPSC lines provide molecular and cellular options to investigate FGFR2 -linked craniosynostosis in the patient-specific genomic context and develop therapeutic modalities.

Refined and benchmarked homemade media for cost-effective, weekend-free human pluripotent stem cell culture L. Truszkowski et al. Open Research Europe 2025 Sep

Abstract

Cost-effective, practical, and reproducible culture of human pluripotent stem cells (hPSCs) is required for basic and translational research. Basal 8 (B8) has emerged as a cost-effective solution for weekend-free and chemically-defined hPSC culture. However, the requirement to home-produce some recombinant growth factors for B8 can hinder access and reproducibility. Moreover, we found the published B8 formulation suboptimal in widely-used normoxic hPSC culture. Lastly, the performance of B8 in functional applications such as genome editing or organoid differentiation required systematic evaluation. We formulated B8 with commercially available, growth factors and adjusted its composition to support normoxic culture of WTC11 human induced pluripotent stem cell line. We compared this formulation (B8+) with commercial Essential 8 (cE8) and a home-made, weekend-free E8 formulation (hE8). We measured pluripotency marker expression and cell cycle by flow cytometry, and investigated the transcriptional profiles by bulk and single-cell RNA sequencing. We further assessed genomic stability, genome editing efficiency, single-cell cloning, and differentiation in both monolayer and organoids. Finally, we validated key findings using male (H1) and female (H9) human embryonic stem cells. hE8 performed comparably to cE8 across most functional assays and cell lines. In contrast, cells in B8+ displayed higher NANOG expression and improved genome editing efficiency. At the same time, B8+ led to gene expression changes indicative of marked lineage priming, reflected in altered morphology and differential response to some differentiation protocols. Both weekend-free media resulted in a modest transcriptional shift towards a less metabolically active state, consistent with intermittent media starvation. Homemade weekend-free media can provide a cost-effective alternative to commercial formulations. hE8, integrating some features of B8 while resembling cE8, emerges as a robust and practical option with limited compromises. B8+, though advantageous in some contexts, warrants caution due to lineage priming effects that may impact differentiation outcomes.

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Related Products

Quality Statement:

For related products, including specialized media, matrices, antibodies, cytokines, and small molecules, visit www.stemcell.com/hPSCworkflow, or contact us at techsupport@stemcell.com.

Safety Statement:

CA WARNING: This product can expose you to chemicals including Nickel Compounds which are known to the State of California to cause cancer and birth defects or other reproductive harm. For more information go to

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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

Product Advantages

What's Included

Request Pricing

Overview

More Information

Protocols and Documentation

Applications

Resources and Publications

Educational Materials (32)

Publications (37)

Abstract

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Abstract

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