CloneR™

Defined, serum-free supplement for the increased cloning efficiency and single-cell survival of human ES and iPS cells.

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CloneR™

Defined, serum-free supplement for the increased cloning efficiency and single-cell survival of human ES and iPS cells.

10 mL
Catalog #05888
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Overview

CloneR™ is a defined, serum-free supplement designed to increase the cloning efficiency and single-cell survival of human embryonic stem cells (ES cells) and induced pluripotent stem cells (iPS cells). CloneR™ enables the robust generation of clonal cell lines without single-cell adaptation, thus minimizing the risk of acquiring genetic abnormalities.

CloneR™ is compatible with the TeSR™ family of media for human ES and iPS cell maintenance as well as your choice of cell culture matrix.
Advantages:
• Greatly facilitates the process of genome editing of human ES and iPS cells
• Compatible with any TeSR™ maintenance medium and your choice of cell culture matrix
• Does not require adaptation to single-cell passaging
• Increases single-cell survival at clonal density across multiple human ES and iPS cell lines
Subtype:
Supplements
Cell Type:
Pluripotent Stem Cells
Species:
Human
Application:
Cell Culture
Brand:
CloneR
Area of Interest:
Cell Line Development; Disease Modeling; Stem Cell Biology
Formulation:
Serum-Free; Defined

Technical Resources

Educational Materials

(2)

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

hPSC Single-Cell Cloning Workflow with CloneR™

Figure 1. hPSC Single-Cell Cloning Workflow with CloneR™

On day 0, human pluripotent stem cells (hPSCs) are seeded as single cells at clonal density (e.g. 25 cells/cm2) or sorted at 1 cell per well in 96-well plates in TeSR™ (mTeSR™1 or TeSR™-E8™) medium supplemented with CloneR™. On day 2, the cells are fed with TeSR™ medium containing CloneR™ supplement. From day 4, cells are maintained in TeSR™ medium without CloneR™. Colonies are ready to be picked between days 10 - 14. Clonal cell lines can be maintained long-term in TeSR™ medium.

CloneR™ Increases the Cloning Efficiency of hPSCs and is Compatible with Multiple hPSC Lines and Seeding Protocols

Figure 2. CloneR™ Increases the Cloning Efficiency of hPSCs and is Compatible with Multiple hPSC Lines and Seeding Protocols

TeSR™ medium supplemented with CloneR™ increases hPSC cloning efficiency compared with cells plated in TeSR™ containing ROCK inhibitor. Cells were seeded (A) at clonal density (25 cells/cm2) in mTeSR™1 and TeSR™-E8™ and (B) by single-cell deposition using FACS (seeded at 1 cell/well) in mTeSR™1.

CloneR™ Increases the Cloning Efficiency of hPSCs at Low Seeding Densities

Figure 3. CloneR™ Increases the Cloning Efficiency of hPSCs at Low Seeding Densities

hPSCs plated in mTeSR™1 supplemented with CloneR™ demonstrated significantly increased cloning efficiencies compared to cells plated in mTeSR™1 containing ROCK inhibitor (10μM Y-27632). Shown are representative images of alkaline phosphatase-stained colonies at day 7 in individual wells of a 12-well plate. H1 human embryonic stem (hES) cells were seeded at clonal density (100 cells/well, 25 cells/cm2) in mTeSR™1 supplemented with (A) ROCK inhibitor or (B) CloneR™ on Vitronectin XF™ cell culture matrix.

CloneR™ Yields Larger Single-Cell Derived Colonies

Figure 4. CloneR™ Yields Larger Single-Cell Derived Colonies

hPSCs seeded in mTeSR™1 supplemented with CloneR™ result in larger colonies than cells seeded in mTeSR™1 containing ROCK inhibitor (10μM Y-27632). Shown are representative images of hPSC clones established after 7 days of culture in mTeSR™1 supplemented with (A) ROCK inhibitor or (B) CloneR™.

Clonal Cell Lines Established Using CloneR™ Display Characteristic hPSC Morphology

Figure 5. Clonal Cell Lines Established Using CloneR™ Display Characteristic hPSC Morphology

Clonal cell lines established using mTeSR™1 or TeSR™-E8™ medium supplemented with CloneR™ retain the prominent nucleoli and high nuclear-to-cytoplasmic ratio characteristic of hPSCs. Representative images at passage 7 after cloning are shown for clones derived from the parental (A) H1 hES cell and (B) WLS-1C human induced pluripotent stem (iPS) cell lines.

Clonal Cell Lines Established with CloneR™ Express High Levels of Undifferentiated Cell Markers

Figure 6. Clonal Cell Lines Established with CloneR™ Express High Levels of Undifferentiated Cell Markers

hPSC clonal cell lines established using mTeSR™1 supplemented with CloneR™ express comparable levels of undifferentiated cell markers, OCT4 (Catalog #60093) and TRA-1-60 (Catalog #60064), as the parental cell lines. (A) Clonal cell lines established from parental H1 hES cell line. (B) Clonal cell lines established from parental WLS-1C hiPS cell line. Data is presented between passages 5 - 7 after cloning and is shown as mean ± SEM; n = 2.

Clonal Cell Lines Established Using CloneR™ Display a Normal Karyotype

Figure 7. Clonal Cell Lines Established Using CloneR™ Display a Normal Karyotype

Representative karyograms of clones derived from parental (A) H1 hES cell and (B) WLS-1C hiPS cell lines demonstrate that the clonal lines established with CloneR™ have a normal karyotype. Cells were karyotyped 5 passages after cloning, with an overall passage number of 45 and 39, respectively.

Clonal Cell Lines Established Using CloneR™ Display Normal Growth Rates

Figure 8. Clonal Cell Lines Established Using CloneR™ Display Normal Growth Rates

Fold expansion of clonal cell lines display similar growth rates to parental cell lines. Shown are clones (red) and parental cell lines (gray) for (A) H1 hES cell and (B) WLS-1C hiPS cell lines.

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.
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