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

Protocols and Documentation

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

Document Type

Product Name

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Lot #

Language

Document Type

Product Name

Catalog #

05230

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All

Language

English

Document Type

Product Name

Catalog #

05230

Lot #

All

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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

Cell Engineering

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

YAP-TEAD regulates the super-enhancer network to control early surface ectoderm commitment Z. Wang et al. Nucleic Acids Research 2026 Jan

Abstract

Super enhancers (SEs), characterized by clusters of enhancers, are instrumental in shaping cellular identity and function. Given this crucial involvement of SEs in cell lineage commitment, and considering the pivotal position of surface ectoderm in differentiating into a wide array of cell types, the study of these SEs holds immense promise for advancing cell-based therapeutic applications. In this study, we profiled the SE landscape in surface ectoderm cells derived from pluripotent stem cell differentiation. By leveraging 3D genomic data, we discerned active histone modifications and frequent chromatin interactions of SEs with target genes. Notably, perturbing specific SE using a CRISPR-dCas9-mediated approach resulted in decreased expression of the connected gene. Subsequently, we constructed a regulatory network of core transcription factors (TFs) operating on SEs and uncovered their control over the differentiation process by forming regulatory network with key TFs, such as TEAD1. Knocking down TEADs attenuated the differentiation process and target gene activation, whereas YAP-TEAD activation expedited the differentiation process by promoting the early establishment of SEs. Collectively, our findings shed light on the crucial role of SEs and identify YAP-TEAD as vital regulators controlling surface ectoderm commitment, thereby providing a novel insight into lineage commitment and stem cell-based epithelial regeneration.

Nanopatterned bioresorbable elastomeric scaffolds to promote neural, glial, and endothelial differentiation using human embryonic and induced pluripotent stem cells I. Romayor et al. Journal of Tissue Engineering 2026 Feb

Abstract

Bioresorbable nanopatterned scaffolds functionalized with polydopamine (PDA) and graphene oxide (GO) have been shown to promote the differentiation of murine neural stem cells (mNSCs) toward neural and glial lineages. Herein, we aim to evaluate the compatibility of these scaffolds for the culture and differentiation of both human embryonic (hESCs) and induced pluripotent (hiPSCs) stem cells. Our results indicate that PDA and GO scaffolds support the topographic alignment of hESCs and hiPSCs cultures, while preserving their pluripotency characteristics. Upon differentiation, PDA and GO scaffolds guide cell specification toward the neuroectoderm germ layer and the neural crest. This promotes enhanced differentiation into both neural and supportive glial cells of the central nervous system (CNS), as well as Schwann cells of the peripheral nervous system (PNS). Moreover, nanopatterned scaffolds also support the differentiation of hESCs and hiPSCs toward endothelial precursors. These findings establish a novel culture platform that enables combined differentiation pathways, potentially relevant for applications in personalized medicine and regenerative cell therapy.

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.

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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

Protocols and Documentation

Applications

Resources and Publications

Educational Materials (32)

Publications (39)

Abstract

Abstract

Abstract

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