Media Additives for hPSC Workflows
The TeSR™ family of feeder-free maintenance media is optimized to support and maintain high-quality human pluripotent stem cell (hPSC) cultures, including embryonic stem (ES) and induced pluripotent stem (iPS) cells. In general, our TeSR™ media is a complete media, consisting of basal and supplement components, tailored for different needs. If single-cell passaging is required for routine hPSC maintenance and expansion, Y-27632 (ROCK inhibitor) or CloneR™2 are added to eTeSR™ (Catalog #100-1215) to enhance cell survival and attachment. Refer to the eTeSR™ Technical Manual for media preparation protocols. Our guidance is to keep the media simple. Hence, it is not necessary to add antibiotics to our TeSR™ media as it may place a selective pressure for a cell line adapted to grow well only when those antibiotics are present in the medium and suppress the detection of culture contamination.
However, there are situations where hPSCs are subjected to stressful conditions in which media additives are required and recommended to improve cell survival and assay performance. STEMCELL Technologies offers several cell culture supplements designed for specific cell applications, such as hPSC single-cell cloning workflows, single-cell sorting, thawing, passaging sensitive cell lines, and plating single cells. This Technical Tip provides a quick comparison chart highlighting several supplements and their recommended applications to help you select the right one for your work.
CloneR™2
Advantages:
- Defined, serum-free TeSR™ supplement
- Compared to CloneR™, it supports higher cloning efficiencies along with generating clones sooner and larger clones after fewer days in culture
- Increased plating efficiency at all densities and after high stress events such as thawing or electroporation
- No single-cell passage adaptation phase required
Recommended Applications:
- Higher cloning efficiency compared to CloneR™. Generating clonal hPSC lines following gene editing and sub-cloning
- Plating hPSCs as single cells for differentiation
- Single-cell sorting using a fluorescence-activated cell sorter (FACS)
- Improving survival after stressful treatments such as electroporation and thawing cryopreserved hPSCs
- Thawing or passaging hPSC aggregates in more sensitive cell lines
- Passaging hPSC cultures as single cells (ensure to test for expected karyotype)
CloneR™
Advantages:
- Defined, serum-free TeSR™ supplement
- Facilitates the process of genome editing of human ES and iPS cells
- No single-cell passage adaptation phase required
Recommended Applications:
- Enables the robust generation of clonal cell lines without single-cell adaptation, thus minimizing the risk of acquiring genetic abnormalities
- Use in clonal and low-density seeding conditions
- Single-cell sorting using a fluorescence-activated cell sorter (FACS)
- Improves embryoid body formation in AggreWell™ protocol
Y-27632 (ROCK inhibitor)
Advantages:
- Highly potent small molecule and selective inhibitor of Rho-associated, coiled-coil containing protein kinase (ROCK)
- Enhances survival of hPSC cells when they are dissociated to single cells by preventing dissociation-induced apoptosis
- Increases cloning efficiency
Recommended Applications:
- Passaging hPSC cultures as single cells (ensure to test for expected karyotype)
- Plating hPSCs as single cells for downstream differentiation
- Improves embryoid body formation in AggreWell™ protocol
- Increases the survival of cryopreserved single hES cells post thaw
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