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mTeSR™ Plus

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

Need a high-quality cell source? Choose from our hiPSC healthy control lines, manufactured with mTeSR™ Plus.

mTeSR™ Plus

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

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


  • Enhanced buffering and stabilized FGF2 support cell quality while allowing for alternate feeding schedules

  • Supports superior culture morphology and cell growth characteristics

  • Enables heightened single-cell survival when used with CloneR™

  • Fully compatible with established genome editing and differentiation protocols

What's Included

  • mTeSR™ Plus Kit, cGMP (Catalog #100-0276)
    • mTeSR™ Plus Basal Medium, 400 mL
    • mTeSR™ Plus 5X Supplement, 100 mL
  • mTeSR™ Plus Kit (Catalog #(100-1130)
    • mTeSR™ Plus Basal Medium, 800 mL
    • mTeSR™ Plus 5x Supplement, 2 x 100 mL

What Our Scientist Says

Innovation is a core value at ƽ. The evolution to mTeSR™ Plus reflects that and our desire to serve the next generation of pluripotent stem cell researchers.

Matthew WongScientist
Matthew Wong, Scientist

Overview

Enjoy weekend-free schedules and enhanced growth characteristics while maintaining cell quality with this stabilized feeder-free maintenance medium for human pluripotent stem cells (hPSCs).

Manufactured under relevant cGMPs, mTeSR™ Plus ensures the highest quality and consistency for fundamental research as well as for cell therapy and investigational new drug research applications. It is based on ձ𳧸™1 (Catalog #85850), the most widely published feeder-free cell culture medium for hPSCs. With stabilized critical medium components, including FGF2, and enhanced pH buffering, you can use mTeSR™ Plus to maintain cell-quality attributes and increase cell expansion rates with either daily or restricted feeding. Each lot of mTeSR™ Plus 5X Supplement is used to prepare complete mTeSR™ Plus medium and then performance-tested in a culture assay using human pluripotent stem cells (hPSCs).

mTeSR™ Plus is compatible with a variety of culture matrices, including Corning® Matrigel® hESC-Qualified Matrix and Vitronectin XF™ (Catalog #07180, developed and manufactured by Nucleus Biologics).

For additional quality information, visit www.stemcell.com/compliance.

To request a Letter of Authorization (LOA) for the FDA Master File for mTeSR™ Plus, click here.
Subtype
Specialized Media
Cell Type
Pluripotent Stem Cells
Species
Human
Application
Cell Culture, Expansion, Maintenance
Brand
TeSR
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Stem Cell Biology
Formulation Category
Serum-Free

Data Figures

Figure 1. mTeSR™ Plus Maintains Optimal pH Levels Throughout a Weekend-Free Protocol

The pH of spent medium from hPSCs cultured in mTeSR™ Plus is higher than that of hPSCs cultured in ձ𳧸™1 and other flexible-feeding medium at similar cell densities. pH and cell numbers were measured after a 72-hour period without feeding. Range of cell numbers shown represent different densities that would be observed throughout a typical passage. This demonstrates that feeds can be skipped for two days at any time during routine maintenance using mTeSR™ Plus while maintaining a pH above 7.0. Note: Cultures were fed double the standard medium volume prior to the 72-hour period without feeds in all media and cell numbers are from one well of a 6-well plate.

Figure 2. mTeSR™ Plus Maintains Consistent Levels of FGF2 Throughout a Weekend-Free Protocol

FGF2 levels remain high in mTeSR™ Plus when cultured at 37°C over a 72 hour time period. Measured by ELISA.

Figure 3. mTeSR™ Plus Supports Higher Cell Numbers

Growth curves were obtained for human ES (H9) cells cultured in ձ𳧸™1 or mTeSR™ Plus on Corning® Matrigel® matrix over 7 days with either daily feeds or restricted feeds. Growth curves were determined by seeding 20,000 cells per well of a 6-well plate as aggregates and counting the cell numbers each day in duplicate wells.

Figure 4. Larger Colonies are Observed in mTeSR™ Plus Cultures

The average colony size per passage (± SEM) was obtained for human ES (H1, H9) and iPS (STiPS-M001, WLS-1C) cells cultured in ձ𳧸™1 (daily feeds) or mTeSR™ Plus (restricted feeds) on Corning® Matrigel® over 10 passages. Size was determined by measuring representative colony diameters at harvest. Note that this data is representative of cultures passaged at a 7-day passaging interval; smaller colony size should be expected if using shorter passaging intervals.

Figure 5. Normal human ES and iPS Cell Morphology is Observed in mTeSR™ Plus Cultures

Images depict undifferentiated human hES (H1) and iPS (WLS-1C) cells cultured on Corning®️ Matrigel®️ matrix in ձ𳧸™1 with daily feeds or mTeSR™ Plus with restricted feeds. Cells retain the prominent nucleoli and high nuclear-to-cytoplasmic ratio characteristic of this cell type after 10 passages. Densely packed cells and multi-layering are prominent when cells are ready to be passaged.

Figure 6. Cells Cultured in mTeSR™ Plus Medium with Restricted Feeding Express Undifferentiated Cell Markers

Human ES (H1, H9) and iPS (WLS-1C, STiPS-M001) cells were characterized using flow cytometry for undifferentiated cell markers, (A) OCT3/4 and (B) TRA-1-60. Graphs show average expression (± SEM) results from analyses of duplicate wells every 5 passages for up to 10-15 passages in ձ𳧸™1 (daily feeds), or mTeSR™ Plus (restricted feeds).

Figure 7. Cells Maintained in mTeSR™ Plus with Restricted Feeding Have Comparable Differentiation Efficiencies to Cells Maintained in ձ𳧸™1

Human ES (H1, H9) and iPS (WLS-1C, STiPS-M001) cells were maintained in ձ𳧸™1 (daily feeds) or mTeSR™ Plus (restricted feeds). Cells were differentiated using directed differentiation protocols and subjected to flow cytometry analysis. Graphs show average expression (± SEM) results from the 4 cell lines. The markers used for flow cytometry for each germ layer are listed in the bar titles.

Figure 8. hPSCs Cultured in mTeSR™ Plus with Restricted Feeding Maintain a Normal Karyotype

Karyograms of (A) human ES (H1) and (B) iPS (WLS-1C) cells cultured in mTeSR™ Plus for 30 passages shows a normal karyotype is retained.

Figure 9. High Cloning Efficiency of hPSCs in mTeSR™ Plus Supplemented with CloneR™

hPSCs (H1, H9, WLS-1C, and STiPS-M001) plated in mTeSR™ Plus with CloneR™ demonstrate cloning efficiencies equal to or greater than hPSCs in ձ𳧸™1 with CloneR™. Cells were seeded at clonal density (25 cells/cm²) in ձ𳧸™1 or mTeSR™ Plus on CellAdhere™ Vitronectin™ XF™-coated plates. n ≧ 3 biological replicates.

Cell morphology images of ES cells plated in ձ𳧸™1 and mTeSR™ Plus and supplemented with CloneR™ immediately following RNP electroporation.

Figure 10. Representative Cell Morphology 24 Hours After RNP Electroporation in ձ𳧸™1 and mTeSR™ Plus

H1-eGFP ES cells were plated in (A) ձ𳧸™1 and (B) mTeSR™ Plus and supplemented with CloneR™ immediately following RNP electroporation. Images were taken 24 hours after electroporation.

Cell images of human ES colonies plated in ձ𳧸™1 and mTeSR™ Plus and supplemented with CloneR™ on CellAdhere™ Vitronectin™ XF™-coated plates.

Figure 11. Clones Derived in mTeSR™ Plus are Larger and Ready to Be Picked at an Earlier Timepoint

Representative images of human ES (H9) colonies taken 8 days following singlecell plating at clonal density (25 cells/cm²) in either (A) ձ𳧸™1 or (B) mTeSR™ Plus supplemented with CloneR™ on CellAdhere™ Vitronectin™ XF™-coated plates.

Cell morphology images of neural progenitor cells maintained in ձ𳧸™1 or mTeSR™ Plus. Arrowheads point to clearly displayed neural rosettes after replating embryoid bodies.

Figure 12. Generation of Neural Progenitor Cells from hPSCs Maintained in mTeSR™ Plus

Human ES (H9) and iPS (STiPS-M001) cells were maintained in (A) ձ𳧸™1 with daily feeds or (B) mTeSR™ Plus with restricted feeds and differentiated using an embryoid body (EB)-based protocol with STEMdiff™ SMADi Neural Induction Kit. Neural progenitor cells derived from hPSCs maintained in either ձ𳧸™1 or mTeSR™ Plus clearly display neural rosettes (arrowheads) after replating EBs.

Immunocytochemistry image of a cerebral organoid cultured in mTeSR™ Plus and directed to cerebral organoids using the STEMdiff™ Cerebral Organoid Kit.

Figure 13. Generation of Cerebral Organoids from hPSCs Maintained in mTeSR™ Plus

Human ES (H9) cells were cultured with mTeSR™ Plus and directed to cerebral organoids using the STEMdiff™ Cerebral Organoid Kit. Image shows apical progenitor marker SOX2 (purple) and neuronal marker TBR1 (green).

Density plots showing CD34+ and CD45+ expression and percentage of cells co-expressing CD34+ and CD45+ and graphs showing total number of viable cells harvested.

Figure 14. Generation of Hematopoietic Progenitor Cells from hPSCs Maintained in mTeSR™ Plus

Human ES (H1, H9) and iPS (STiPS-M001, WLS-1C) cell lines maintained in ձ𳧸™1 (daily feeds) or mTeSR™ Plus (restricted feeds) were differentiated to hematopoietic progenitor cells using the STEMdiff™ Hematopoietic Kit. At the end of the differentiation period, cells were harvested from the supernatant and analyzed by flow cytometry for co-expression of CD34+ and CD45+ . (A) Representative density plots showing CD34+ and CD45+ expression, (B) percentage of cells co-expressing CD34+ and CD45+ , and (C) total number of viable cells harvested are shown. Data are expressed as the mean (± SEM); n=4.

Microelectrode array and flow cytometry of human ES and iPS cells maintained in ձ𳧸™1 (daily feeds) or mTeSR™ Plus (restricted feeds) and differentiated to cardiomyocytes using the STEMdiff™ Cardiomyocyte Differentiation Kit.

Figure 15. Generation of Cardiomyocytes from hPSCs Maintained in mTeSR™ Plus

Human ES (H9) and iPS (WLS-1C) cells were maintained in ձ𳧸™1 (daily feeds) or mTeSR™ Plus (restricted feeds) and differentiated to cardiomyocytes using the STEMdiff™ Cardiomyocyte Differentiation Kit. At the end of the differentiation period, cells were harvested and analyzed by microelectrode array (MEA) and flow cytometry. (A) Representative MEA voltage recordings of cardiomyocytes (day 20) demonstrate a characteristic electrical profile and stable beat rate. (B) Percentages of cells expressing cTNT and (C) total number of viable cells harvested are shown. Data are expressed as the mean (± SEM); n=2.

Immunocytochemistry image of an intestinal organoid cultured in mTeSR™ Plus and directed to intestinal organoids using the STEMdiff™ Intestinal Organoid Kit.

Figure 16. Generation of Intestinal Organoids from hPSCs Maintained in mTeSR™ Plus

Human ES (H9) cells were cultured with mTeSR™ Plus and directed to intestinal organoids using the STEMdiff™ Intestinal Organoid Kit. Image shows markers of the intestinal epithelium EpCAM (green) and CDX2 (red), and intestinal mesenchyme marker vimentin (white). Nuclei are counterstained with DAPI (blue).

Density plots and quantitative analysis showing CXCR4 and SOX17 expression in cells cultured in ձ𳧸™1 (daily feeds) or mTeSR™ Plus (restricted feeds), following 5 days of differentiation using the STEMdiff™ Definitive Endoderm Kit.

Figure 17. Generation of Definitive Endoderm from hPSCs Maintained in mTeSR™ Plus

(A) Representative density plots showing CXCR4 and SOX17 expression in cells cultured in ձ𳧸™1 (daily feeds) or mTeSR™ Plus (restricted feeds), following 5 days of differentiation using the STEMdiff™ Definitive Endoderm Kit. (B) Quantitative analysis of definitive endoderm formation in multiple hPSC lines (H9, STiPS-M001, WLS-1C) maintained with ձ𳧸™1 or mTeSR™ Plus as measured by co-expression of CXCR4 and SOX17. Data are expressed as the mean percentage of cells (± SEM) expressing both markers; n=3.

Density plots and quantitative analysis showing PDX-1 and NKX6.1 expression in cells cultured in ձ𳧸™1 or mTeSR™ Plus, following 5 days of differentiation using the STEMdiff™ Pancreatic Progenitor Kit.

Figure 18. Generation of Pancreatic Progenitors from hPSCs Maintained in mTeSR™ Plus

(A) Representative density plots showing PDX-1 and NKX6.1 expression in cells cultured in ձ𳧸™1 (daily feeds) or mTeSR™ Plus (restricted feeds), following differentiation using the STEMdiff™ Pancreatic Progenitor Kit. (B) Quantitative analysis of pancreatic progenitor formation in multiple hPS (H9, STiPS-M001, WLS-1C) cell lines maintained with ձ𳧸™1 or mTeSR™ Plus as measured by co-expression of PDX-1 and NKX6.1. Data are expressed as the mean percentage of cells (± SEM) expressing both markers; n=3.

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 #
100-0276, 100-1130
Lot #
All
Language
English
Document Type
Product Name
Catalog #
100-0276, 100-1130
Lot #
All
Language
English
Document Type
Product Name
Catalog #
100-0276, 100-1130
Lot #
All
Language
English
Document Type
Product Name
Catalog #
100-0276, 100-1130
Lot #
All
Language
English
Document Type
Product Name
Catalog #
100-1130
Lot #
All
Language
English
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Applications

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Resources and Publications

Educational Materials (56)

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Publications (271)

Exceptional Longevity Modifying Allele APOE2 Promotes DNA Signaling Pathways Resisting Cellular Senescence in Human Neurons C. Gerónimo‐Olvera et al. Aging Cell 2026 May

Abstract

Genome‐wide association studies (GWAS) have identified APOE2 allele as linked to exceptional longevity, with carriers exhibiting a reduced risk of Alzheimer's disease (AD). Apolipoprotein E (APOE), a glycoprotein involved in lipid transport, has three major alleles. However, alterations in lipid metabolism alone do not fully explain APOE2's protective effects. In contrast, APOE4 is the strongest genetic risk factor for AD. To investigate how APOE2 promotes neuronal longevity and confers neuroprotection, we generated human isogenic APOE iPSC‐derived models of both inhibitory GABAergic and excitatory neurons. In GABAergic neurons, APOE alleles differentially influenced endogenous DNA damage, DNA repair, and neuronal motility. Single‐cell RNA sequencing revealed APOE4‐specific gene expression signatures associated with AD, whereas APOE2 GABAergic neurons were enriched for DNA repair and signaling pathways. Consistent with this, APOE2 neurons exhibited significantly lower levels of DNA damage. APOE4 GABAergic neurons exhibit increased expression of repetitive ribosomal RNA, which is associated with DNA damage and cellular senescence. To determine whether the effects extended to excitatory neurons, we used a separate human model of Ngn2‐induced glutamatergic neurons, and found that APOE2 excitatory neurons were more resistant to cellular senescence and DNA damage than isogenic APOE3 and APOE4 neurons. Similarly, we found human APOE2‐targeted replacement mice exhibited less nucleolar enlargement and increased nuclear Lamin A/C, Hmgb1, and H3K9me3 compared to APOE4 counterparts. Together, our findings identify DNA repair and suppression of senescence‐associated processes as key mechanisms by which APOE2 is associated with neuronal resilience, providing mechanistic insight into its association with exceptional longevity and protection against AD. Neurons expressing APOE2 were more resistant to endogenous DNA damage, activated transcriptional signaling pathways associated with DNA repair, and were resilient to stress‐induced DNA damage and cellular senescence. In contrast, APOE4 neurons exhibited elevated expression of rRNA repetitive elements and were prone to becoming senescent.
Human iPSC-Derived Blood Vessel Organoids for Studying Chronic Hypoxia-Induced Microvascular Dysfunction P. S. Martinez et al. Journal of Histochemistry and Cytochemistry 2026 May

Abstract

Microvascular dysfunction due to hypoxia is a key contributor in the pathogenesis of many disorders including cancer and retinal and cardiovascular diseases, but relevant human models are missing. Here, we present a robust 3D in vitro method with the use of human induced pluripotent stem cell–derived blood vessel organoids to analyze in vitro microvascular remodeling. We present a detailed practical pipeline combining optical tissue clearing, high-resolution immunofluorescence, and surface marker analysis to quantitatively assess hypoxia-driven changes in endothelial cells, pericytes, and the basal lamina. Exposure of these blood vessel organoids to chronic hypoxia (1% O2) for 1 week recapitulated key pathological features, including structural remodeling and a dysregulated secretome with altered vascular endothelial growth factor signaling. This approach establishes a versatile and human-relevant platform to study microvascular remodeling induced by chronic hypoxia and other pathological stimuli and their contribution to microvascular-related diseases.
Generation of Functional Patient-Specific Thymus Organoids From Human Pluripotent Stem Cells (hPSCs) Using Air–Liquid Interface Culture S. A. Ramos, H. A. Russ Bio-protocol 2026 May

Abstract

The thymus is critical for the establishment of a functional and self-tolerant adaptive immune system, but it involutes with age, resulting in reduced naive T-cell output. Generation of a functional human thymus from human pluripotent stem cells (hPSCs) is an attractive regenerative medicine strategy. Direct differentiation of thymic epithelial progenitors (TEPs) from hPSCs has been demonstrated in vitro, but functional thymic epithelial cells (TECs) develop only after transplantation of TEPs in vivo. Functional human reaggregated thymic organoid cultures (RTOCs) and artificial thymic organoids (ATOs) cultured at the air–liquid interface support T-cell development in vitro and in vivo and permit the interrogation of human thymic function and T-cell development. However, these approaches require access to primary human tissues or murine bone marrow stromal cells, are allogeneic, and do not support negative selection. Recently, we reported the directed differentiation of induced PSCs (iPSCs) to functional thymic epithelial progenitors (TEPs) that support murine T-cell development after transplantation in nude mice. Here, we combined hPSC-derived TEPs, hematopoietic progenitor cells (HPCs), and mesenchymal cells, differentiated from the same hPSC line, and generated functional isogenic stem cell–derived thymic organoids (sTOs). Our revised protocol improves our TEP differentiation process and allows the generation of functional isogenic, patient-specific thymic organoids in vitro.
Need a high-quality cell source? Choose from our hiPSC healthy control lines, manufactured with mTeSR™ Plus.