Description
The NanoSparkTM STEM-T Soluble T Cell Activator is a culture-ready reagent for in vitro activation and expansion of human T cells. The nanoscale of the NanoSparkTM STEM-T protein complex allows for sterile filtration and removal of excess reagent via either media exchange or simple centrifugal cell washing.
NanoSparkTM STEM-T: High Stemlike T cell Production
NanoSparkTM STEM-T is best in class, particularly for preserving stemlike CD8+ T cells important for higher efficacy. Competing products introduce clinical risks to the process by using synthetic ingredients that may contaminate T cell therapy if not painstakingly removed from the expanded culture.
The NanoSparkTM product line is modular and plug-and-play by design, using a proprietary self-assembly protein nanoparticle complex that is inherently soluble and can be filter sterilized.
How Does the NanoSparkTM STEM-T Soluble T Cell Activator Work?
The NanoSparkTM STEM-T Soluble T Cell Activator contains self-assembling protein nanoparticles with functional antibodies bound to the surface through high-affinity non-covalent interactions. These anti-CD3 and anti-CD28 antibodies are arranged in a proprietary biophysical combination on the NanoSparkTM platform to drive best-in-class performance that favors stemlike CD8+ T cell production.
The NanoSparkTM STEM-T Soluble T Cell Activator activates resting human CD3+ T cells originating from purified T lymphocytes or patient peripheral blood mononuclear cells (PBMCs).
Inquire About Nanotein’s T Cell Activator
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Convenient
Soluble, magnetic bead-free, protein-based, and ready-to-use.
High Performance
Excellent total T cell expansion with industry-leading clinically desirable CD8+ stem cell memory T cell expansion.
Reliable
Employs anti-CD3/CD28 antibodies and maintains high cell viability.
Documentation & Protocols
Consult the data sheets and supporting documentation for usage instructions and additional details, or explore the recommended protocols, which include suggested catalog numbers for cytokine supplements and media.
Data Sheets
These protocols are recommended and include suggested catalog numbers of cytokine supplements and media to use.
The NanoSparkTM STEM-T Soluble T Cell Activator is used in vitro to induce activation and CAR gene delivery uptake in stemlike CD8+ cells via anti-CD3/CD28 pathways. It is intended for ex vivo activation and expansion of CD3+ T Lymphocytes (human resting T cells) from T cell isolates or peripheral blood mononuclear cells (PBMCs). The activator should be stored at -80 °C long-term. Once thawed, store at 4 °C protected from light for up to one month.
The NanoSparkTM STEM-T Soluble T Cell Activator is designed for use with a cytokine-supplemented T cell expansion medium. Suggested protocols have been developed for common cytokine-supplemented strategies. Optimization of the provided general protocol information may be necessary depending on your experimental objectives.
Standard STEM-T Expansion Protocol & Data Sheet
Short 3-Day STEM-T Expansion Protocol & Data Sheet
Certificate of Analysis (CoA)
Lot-specific STEM-T CoA will be shipped with the product. Contact customer service if additional copies are required.
Nanotein Example STEM-T CoA
Safety Data Sheet (SDS)
Carefully read product SDS before use.
NanosparkTM STEM-T SDS
Tips for Best Results Using the NanoSparkTM STEM-T Soluble T Cell Activator
- Seed CD3+ T cells/PBMCs at 7 x 105 cells/cm2 in culture media.
- Add volume of NanoSparkTM STEM-T Soluble T Cell Activator as recommended in table above to achieve specific goals.
- Incubate cells at 37 °C and 5% CO2 in a humidified incubator.
- For cryopreserved cells, gently thaw cells and exchange them into culture media before starting.
- For optimal cell expansion and maintenance, ensure the activator is in culture media (conditioned or fresh) for at least 72 hours (removal is unnecessary for expansion).
- Be sure to add fresh cytokine-supplemented culture media every two to three days as needed.
If using transduction (optional), wait 24-72 hours after adding NanoSparkTM STEM-T Soluble T Cell Activator to the cell culture before adding the viral vector. If using electroporation, then electroporate 72 hours after activator addition.
Data & Research
Enhanced T Cell Expansion and Viability with NanoSparkTM STEM-T Soluble T-Cell Activator
A) Fold Change of CD3+ Cells, B) % of CD4+ & CD8+ Cells, C) CD4+ & CD8+ T-Cell Phenotype Percentage. NanoSparkTM STEM-T Soluble T-Cell Activator was cultured in StemCell’s ImmunoCult-XF T Cell Expansion Medium (xeno-free) supplemented with IL-7 and IL-15. Cells were expanded for 10 days and analyzed on a flow cytometer on days 0, 7, and 10. Cells were labeled with CD4+, CD8+, CD45RA, CCR7, and CD95 fluorescent antibodies (Schmueck-Henneresse et al 2017). Expansion with NanoSparkTM STEM-T Soluble T-Cell Activator enhances total cell viability and stimulates expansion of CD8+ T-Cells and promotes the stem-like phenotypes of CD4+ and CD8+ T-Cells: Tnaive, Tscm, and Tcm.
Optimized T-Cell Expansion and Phenotypic Development with NanoSparkTM STEM-T in G-Rex6M System
A) Fold Change of CD3+ Cells, B) % of CD4+ & CD8+ Cells, C) CD4+ & CD8+ T-Cell Phenotype Percentage. NanoSparkTM STEM-T Soluble T-Cell Activator was cultured in StemCell’s ImmunoCult-XF T Cell Expansion Medium (xeno-free) supplemented with IL-7 and IL-15. Cells were expanded for 10 days in a G-rex6M and analyzed on a flow cytometer on days 0, and 10. Cells were labeled with CD4+, CD8+, CD45RA, CCR7, and CD95 fluorescent antibodies (Schmueck-Henneresse et al 2017). Expansion with NanoSparkTM STEM-T Soluble T-Cell Activator enhances total cell viability and stimulates expansion of CD8+ T-Cells and promotes the stem-like phenotypes of CD4+ and CD8+ T-Cells: Tnaive, Tscm, and Tcm.
Quality & Safety
Nanotein assessed the NanoSparkTM platform with adherence to rigorous ISO 10993 standards in preparation for GMP manufacturing. The composition of the NanoSparkTM reagent is protein-based, and consequently naturally biodegradable, which provides a significant safety advantage over magnetic bead-based and synthetic products. A reputable medical device CRO was commissioned to complete the in vitro & in vivo ISO 10993 biocompatibility testing. The tests were run at >20X the predicted possible carry-over amount, which resulted in no toxicity or adverse effects:
