mTeSR™ Plus

Stabilized feeder-free maintenance medium for human ES and iPS cells

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mTeSR™ Plus

Stabilized feeder-free maintenance medium for human ES and iPS cells

1 Kit
Catalog #05825
279 USD

Overview

mTeSR™ Plus is a serum-free, stabilized cell culture medium for the feeder-free maintenance and expansion of human embryonic stem (ES) and induced pluripotent stem (iPS) cells. It is based on the mTeSR™1 formulation, the most widely published feeder-free cell culture medium for human ES and iPS cells.

To enhance cell-quality attributes during restricted feeds, critical medium components have been stabilized, including FGF2, and medium pH is more consistent. As a result, mTeSR™ Plus allows for greater cell expansion rates with daily feeding, while also maintaining cell quality during restricted feeding schedules.

mTeSR™ Plus is compatible with a variety of culture matrices, including Corning® Matrigel® hESC-Qualified Matrix (Corning Catalog #354277) and Vitronectin XF™ (Catalog #07180, a matrix developed and manufactured by Primorigen Biosciences).

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).
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
Components:
  • mTeSR™ Plus Basal Medium, 400 mL
  • mTeSR™ Plus 5X Supplement, 100 mL
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:
Serum-Free

Scientific Resources

Educational Materials

(9)

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

Data

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 mTeSR™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 mTeSR™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 mTeSR™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 mTeSR™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 mTeSR™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 mTeSR™1

Human ES (H1, H9) and iPS (WLS-1C, STiPS-M001) cells were maintained in mTeSR™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 mTeSR™1 with CloneR™. Cells were seeded at clonal density (25 cells/cm²) in mTeSR™1 or mTeSR™ Plus on CellAdhere™ Vitronectin™ XF™-coated plates. n ≧ 3 biological replicates.

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.