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Rhil 7

Manufactured by CellGenix
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RhIL-7 is a recombinant human interleukin-7 protein. Interleukin-7 is a cytokine that plays a crucial role in the development and maintenance of T cells.

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Lab products found in correlation

Rhil 15, by CellGenix (3 mentions) Clinimacs device, by Miltenyi Biotec (2 mentions) Knockout sr, by Thermo Fisher Scientific (2 mentions) Rhil 21, by Miltenyi Biotec (2 mentions) Cts dynabeads cd3 cd28, by Thermo Fisher Scientific (2 mentions) Sepax 2rm device, by Cytiva (2 mentions) Cryostor cs5, by BioLife Solutions (2 mentions) X vivo 15, by Lonza (2 mentions) Streptomycin, by Harvard Bioscience (1 mentions) Ifnγ secretion assay cell enrichment and detection kit, by Miltenyi Biotec (1 mentions) Vle rpmi 1640, by PAN Biotech (1 mentions)

3 protocols using rhil 7

1

Anti-B7-H3 CAR-T Cell Production

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CD4+ and CD8+ T cells were isolated from patient apheresis products using the CliniMACS device (Miltenyi Biotec). A 1:1 mixture of CD4-enriched and CD8-enriched cell fractions were then pooled, suspended in X-Vivo 15 (Lonza) media supplemented with 2% KnockOut SR (Life Technologies), 5 ng/mL rhIL-7 (CellGenix), 0.5 ng/mL rhIL-15 (CellGenix) and 10 ng/mL rhIL-21 (Miltenyi Biotec), initiated into culture in a G-Rex 100MCS vessel (Wilson-Wolf), and stimulated using CD3/CD28 CTS® Dynabeads (Life Technologies). Cell products were then transduced with a GMP-grade SIN (self-inactivating) lentivirus encoding the anti-B7-H3CAR, the methotrexate-resistant human DHFR mutein huDHFRdm, and the cell-surface marker EGFRt. On day 3 of culture, 50nM methotrexate (Mylan) was added to the culture vessel to select for cells possessing DHFRdm. On day 7 of culture, CD3/CD28 CTS® beads were removed from the cell suspension using the Dynamag CTS device. Following 10–13 days in culture, patient cells were harvested and washed using the Sepax 2RM device (Cytiva) and resuspended in CryoStor-CS5 (Biolife Solutions) for cryopreservation in CellSeal closed-system vials (Sexton).
Vitanza N.A., Wilson A.L., Huang W., Seidel K., Brown C., Gustafson J.A., Yokoyama J.K., Johnson A.J., Baxter B.A., Koning R.W., Reid A.N., Meechan M., Biery M.C., Myers C., Rawlings-Rhea S.D., Albert C.M., Browd S.R., Hauptman J.S., Lee A., Ojemann J.G., Berens M.E., Dun M.D., Foster J.B., Crotty E.E., Leary S.E., Cole B.L., Perez F.A., Wright J.N., Orentas R.J., Chour T., Newell E.W., Whiteaker J.R., Zhao L., Paulovich A.G., Pinto N., Gust J., Gardner R.A., Jensen M.C, & Park J.R. (2022). Intraventricular B7-H3 CAR T Cells for Diffuse Intrinsic Pontine Glioma: Preliminary First-in-Human Bioactivity and Safety. Cancer Discovery, 13(1), 114-131.
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2

Engineered Antigen-Specific T Cell Therapy

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CD4+ and CD8+ T cells were isolated from patient apheresis products using the CliniMACS device (Miltenyi Biotec). A 1:1 mixture of CD4-enriched and CD8-enriched cell fractions were then pooled, suspended in X-Vivo 15 (Lonza) media supplemented with 2% KnockOut SR (Life Technologies), 5 ng/mL rhIL7 (CellGenix), 0.5 ng/mL rhIL15 (CellGenix), and 10 ng/mL rhIL-21 (Miltenyi Biotec), initiated into culture in a G-Rex 100MCS vessel (Wilson-Wolf), and stimulated using CD3/CD28 CTS Dynabeads (Life Technologies). Cell products were then transduced with a GMP-grade SIN (self-inactivating) lentivirus encoding the anti-B7-H3 CAR, the MTX-resistant human DHFR mutein huDHFRdm, and the cell-surface marker EGFRt. On day 3 of culture, 50 nmol/L MTX (Mylan) was added to the culture vessel to select for cells possessing DHFRdm. On day 7 of culture, CD3/CD28 CTS beads were removed from the cell suspension using the Dynamag CTS device. Following 10 to 13 days in culture, patient cells were harvested and washed using the Sepax 2RM device (Cytiva) and resuspended in CryoStor-CS5 (Biolife Solutions) for cryopreservation in CellSeal closed-system vials (Sexton).
Vitanza N.A., Wilson A.L., Huang W., Seidel K., Brown C., Gustafson J.A., Yokoyama J.K., Johnson A.J., Baxter B.A., Koning R.W., Reid A.N., Meechan M., Biery M.C., Myers C., Rawlings-Rhea S.D., Albert C.M., Browd S.R., Hauptman J.S., Lee A., Ojemann J.G., Berens M.E., Dun M.D., Foster J.B., Crotty E.E., Leary S.E., Cole B.L., Perez F.A., Wright J.N., Orentas R.J., Chour T., Newell E.W., Whiteaker J.R., Zhao L., Paulovich A.G., Pinto N., Gust J., Gardner R.A., Jensen M.C, & Park J.R. (2022). Intraventricular B7-H3 CAR T Cells for Diffuse Intrinsic Pontine Glioma: Preliminary First-in-Human Bioactivity and Safety. Cancer Discovery, 13(1), 114-131.
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3

Isolation and Culture of SARS-CoV-2-Specific T Cells

Cited 1 time
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Virus-specific T cells were isolated from PBMCs derived from 100 mL of peripheral blood from convalescent donors following a 6-h stimulation with overlapping SARS-CoV-2-specific peptide pools (JPT Peptide Technologies; 1 μg/mL each) using an IFN-γ Secretion Assay—Cell Enrichment and Detection Kit according to the manufacturer’s instructions (Miltenyi Biotec). Isolated virus-specific T cells were cultured in complete medium (VLE RPMI 1640 [PAN-Biotech] supplemented with penicillin [100 IU/mL], streptomycin [Biochrom], 10% fetal calf serum [FCS, PAA], 10 ng/mL recombinant human IL-7 [rhIL-7] and rhIL-15 [CellGenix]) in 24-well plates, in humidified incubators at 37°C and 5% CO2 as described previously.88 (link),122 (link) Cells were split 1:1 upon reaching 100% confluency.
Peter L., Wendering D.J., Schlickeiser S., Hoffmann H., Noster R., Wagner D.L., Zarrinrad G., Münch S., Picht S., Schulenberg S., Moradian H., Mashreghi M.F., Klein O., Gossen M., Roch T., Babel N., Reinke P., Volk H.D., Amini L, & Schmueck-Henneresse M. (2022). Tacrolimus-resistant SARS-CoV-2-specific T cell products to prevent and treat severe COVID-19 in immunosuppressed patients. Molecular Therapy. Methods & Clinical Development, 25, 52-73.
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