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KCL-ENG-104
⚠ FOR RESEARCH USE ONLY — Not for diagnostic or therapeutic procedures
KCL-ENG-104 is a stably engineered subline of RPMI-8226, a human B lymphocyte line derived from a multiple myeloma patient. The parental RPMI-8226 line typically expresses plasma cell markers (CD38, CD138) with low to negative CD19. This engineered line was generated by lentiviral transduction with a human CD19 (hCD19) expression cassette and antibiotic selection, resulting in high, stable surface CD19 expression on a suspension-growing B-cell background.
Greater than 95% of cells express CD19 by flow cytometry, with expression stable for a minimum of 10 passages without selection pressure. The line retains the RPMI-8226 plasma cell phenotype (CD38+, CD138+, CD20-) alongside engineered CD19 expression, making it uniquely suited for studies requiring a CD19-positive myeloma background — including CAR-T and CAR-NK cytotoxicity assays, bispecific antibody (BiTE) screening, and ADCC evaluation.
All lots are quality tested prior to release for CD19 expression, post-thaw viability, mycoplasma, sterility, viral contamination, and STR identity.
RPMI-1640 supplemented with 10 to 20% heat-inactivated FBS, 1% Penicillin-Streptomycin, and 2 mM L-Glutamine. Maintain cells at 0.5 to 2×10&sup6; cells/mL. For long-term culture, use intermittent Puromycin selection (1 to 2 μg/mL every 3 to 4 passages) to preserve CD19 expression. Omit selection antibiotics during co-culture assays with immune cells.
All lots are quality tested prior to release. Surface marker expression is confirmed by flow cytometry within 5 passages post-thaw.
| Marker | Result | Typical % | Notes |
|---|---|---|---|
| CD19 (human) | ✓ Positive | >95% | Engineered transgene; CAR-T / bispecific antibody target |
| CD38 | ✓ Positive | >90% | Plasma cell marker; confirms B-cell lineage |
| CD138 (Syndecan-1) | ✓ Positive | >85% | Confirms plasmacytic origin |
| BCMA (TNFRSF17) | ✓ Positive | >80% | Myeloma-specific immunotherapy target |
| HLA-DR | ≈ Variable | ~20–40% | B-cell activation state; user-verifiable |
| CD20 | ✗ Negative | <2% | Distinguishes from mature B-cell lines (e.g., Raji, Daudi) |
| CD3 | ✗ Negative | <1% | Confirms absence of T-cell contamination |
| CD34 | ✗ Negative | <1% | Confirms differentiated B-cell stage |
| Test | Result | Method |
|---|---|---|
| Post-thaw viability | ✓ >90% | Flow cytometry (7-AAD) |
| Mycoplasma | ✗ Negative | qPCR |
| Sterility — Bacteria & Fungi | ✗ Negative | 14-day culture (BHI/TSB media) |
| HIV-1, HIV-2, HBV, HCV, HTLV-1/2 | ✗ Negative | PCR or ELISA |
| Cell line identity (STR profiling) | ✓ Confirmed | Matches RPMI-8226 reference + hCD19 transgene (PCR) |
Certificates of Analysis with lot-specific QC data are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-104 is shipped as a cryopreserved vial on dry ice. These cells grow in suspension — do not expect adherent monolayer formation. Follow the steps below upon receipt.
Maintain at 0.5 to 2×10&sup6; cells/mL. Split 1:2 to 1:4 every 48 to 72 hours. Do not allow density to exceed 2×10&sup6; cells/mL. For long-term culture, apply intermittent Puromycin selection (1 to 2 μg/mL) every 3 to 4 passages to maintain stable CD19 expression. Full protocols are available on the Kulture Scientific protocol page.
KCL-ENG-104 is available as a cryopreserved vial containing at least 5×10&sup5; viable cells in serum-free freezing medium.
| Product | Cat. No. | Format | Cell Number |
|---|---|---|---|
| Human B Lymphocyte (RPMI-8226) / CD19 Positive Stable Cells | KCL-ENG-104 | Cryopreserved Vial | ≥5×105 cells/vial |
- Storage — store in liquid nitrogen vapor phase (−150°C to −196°C); this is the only recommended storage condition for cryopreserved cells
- Do not store at −80°C or −20°C — neither temperature is suitable for long-term cell storage; ice crystal formation at these temperatures will irreversibly damage cell membranes and compromise viability
- Temperature sensitivity — minimize time outside cryogenic storage; cells are sensitive to temperature fluctuations
- Freeze-thaw cycles — repeated freeze-thaw cycles compromise viability and CD19 expression stability; thaw only the quantity needed for immediate use
- Stability — cells stored in liquid nitrogen are stable indefinitely when maintained under correct cryogenic conditions
- Shipping condition — cryopreserved vials are shipped on dry ice; inspect packaging upon receipt and transfer to liquid nitrogen storage immediately
- Freezing medium — cells are cryopreserved in serum-free freezing medium containing DMSO; wear nitrile gloves during thaw procedures and avoid skin contact
- Certificate of Analysis — lot-specific QC documentation including flow cytometry data, viability, sterility, and STR profile available upon request
For research purposes only. Not for use in diagnostic or therapeutic procedures. This product has not been tested or validated for clinical use. Not for direct administration into humans or animals.
This cell line should be handled under Biosafety Level 2 (BSL-2) conditions based on the following characteristics:
- Human-derived origin — all human-derived cell lines require BSL-2 containment under Universal Precautions regardless of donor screening status; derived from a multiple myeloma patient
- Lentiviral transduction history — generated using lentiviral vectors; while replication-incompetent, standard BSL-2 containment for lentivirally transduced lines applies until confirmed negative for replication-competent lentivirus (RCL)
- Bloodborne pathogen risk — human-derived material of hematopoietic origin; treat as potentially infectious for EBV, Hepatitis, and other bloodborne pathogens
Users must consult their institutional biosafety committee (IBC) prior to use. Individual institutions may impose additional requirements based on local policy.
- Human-derived hematopoietic material — treat as potentially infectious; follow BSL-2 guidelines
- Lentiviral transduction history — handle in accordance with IBC guidelines for lentivirally transduced suspension cell lines
- Suspension growth — aerosol risk is elevated with suspension cultures; use appropriate containment when pipetting or centrifuging
- DMSO (cryoprotectant) — skin-penetrating solvent present in freezing medium; wear nitrile gloves at all times during thawing
- Disposal — decontaminate all cell culture waste with 10% bleach prior to disposal in accordance with local biohazardous waste regulations
⚠ FOR RESEARCH USE ONLY — Not for diagnostic or therapeutic procedures
KCL-ENG-104 is a stably engineered subline of RPMI-8226, a human B lymphocyte line derived from a multiple myeloma patient. The parental RPMI-8226 line typically expresses plasma cell markers (CD38, CD138) with low to negative CD19. This engineered line was generated by lentiviral transduction with a human CD19 (hCD19) expression cassette and antibiotic selection, resulting in high, stable surface CD19 expression on a suspension-growing B-cell background.
Greater than 95% of cells express CD19 by flow cytometry, with expression stable for a minimum of 10 passages without selection pressure. The line retains the RPMI-8226 plasma cell phenotype (CD38+, CD138+, CD20-) alongside engineered CD19 expression, making it uniquely suited for studies requiring a CD19-positive myeloma background — including CAR-T and CAR-NK cytotoxicity assays, bispecific antibody (BiTE) screening, and ADCC evaluation.
All lots are quality tested prior to release for CD19 expression, post-thaw viability, mycoplasma, sterility, viral contamination, and STR identity.
RPMI-1640 supplemented with 10 to 20% heat-inactivated FBS, 1% Penicillin-Streptomycin, and 2 mM L-Glutamine. Maintain cells at 0.5 to 2×10&sup6; cells/mL. For long-term culture, use intermittent Puromycin selection (1 to 2 μg/mL every 3 to 4 passages) to preserve CD19 expression. Omit selection antibiotics during co-culture assays with immune cells.
All lots are quality tested prior to release. Surface marker expression is confirmed by flow cytometry within 5 passages post-thaw.
| Marker | Result | Typical % | Notes |
|---|---|---|---|
| CD19 (human) | ✓ Positive | >95% | Engineered transgene; CAR-T / bispecific antibody target |
| CD38 | ✓ Positive | >90% | Plasma cell marker; confirms B-cell lineage |
| CD138 (Syndecan-1) | ✓ Positive | >85% | Confirms plasmacytic origin |
| BCMA (TNFRSF17) | ✓ Positive | >80% | Myeloma-specific immunotherapy target |
| HLA-DR | ≈ Variable | ~20–40% | B-cell activation state; user-verifiable |
| CD20 | ✗ Negative | <2% | Distinguishes from mature B-cell lines (e.g., Raji, Daudi) |
| CD3 | ✗ Negative | <1% | Confirms absence of T-cell contamination |
| CD34 | ✗ Negative | <1% | Confirms differentiated B-cell stage |
| Test | Result | Method |
|---|---|---|
| Post-thaw viability | ✓ >90% | Flow cytometry (7-AAD) |
| Mycoplasma | ✗ Negative | qPCR |
| Sterility — Bacteria & Fungi | ✗ Negative | 14-day culture (BHI/TSB media) |
| HIV-1, HIV-2, HBV, HCV, HTLV-1/2 | ✗ Negative | PCR or ELISA |
| Cell line identity (STR profiling) | ✓ Confirmed | Matches RPMI-8226 reference + hCD19 transgene (PCR) |
Certificates of Analysis with lot-specific QC data are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-104 is shipped as a cryopreserved vial on dry ice. These cells grow in suspension — do not expect adherent monolayer formation. Follow the steps below upon receipt.
Maintain at 0.5 to 2×10&sup6; cells/mL. Split 1:2 to 1:4 every 48 to 72 hours. Do not allow density to exceed 2×10&sup6; cells/mL. For long-term culture, apply intermittent Puromycin selection (1 to 2 μg/mL) every 3 to 4 passages to maintain stable CD19 expression. Full protocols are available on the Kulture Scientific protocol page.
KCL-ENG-104 is available as a cryopreserved vial containing at least 5×10&sup5; viable cells in serum-free freezing medium.
| Product | Cat. No. | Format | Cell Number |
|---|---|---|---|
| Human B Lymphocyte (RPMI-8226) / CD19 Positive Stable Cells | KCL-ENG-104 | Cryopreserved Vial | ≥5×105 cells/vial |
- Storage — store in liquid nitrogen vapor phase (−150°C to −196°C); this is the only recommended storage condition for cryopreserved cells
- Do not store at −80°C or −20°C — neither temperature is suitable for long-term cell storage; ice crystal formation at these temperatures will irreversibly damage cell membranes and compromise viability
- Temperature sensitivity — minimize time outside cryogenic storage; cells are sensitive to temperature fluctuations
- Freeze-thaw cycles — repeated freeze-thaw cycles compromise viability and CD19 expression stability; thaw only the quantity needed for immediate use
- Stability — cells stored in liquid nitrogen are stable indefinitely when maintained under correct cryogenic conditions
- Shipping condition — cryopreserved vials are shipped on dry ice; inspect packaging upon receipt and transfer to liquid nitrogen storage immediately
- Freezing medium — cells are cryopreserved in serum-free freezing medium containing DMSO; wear nitrile gloves during thaw procedures and avoid skin contact
- Certificate of Analysis — lot-specific QC documentation including flow cytometry data, viability, sterility, and STR profile available upon request
For research purposes only. Not for use in diagnostic or therapeutic procedures. This product has not been tested or validated for clinical use. Not for direct administration into humans or animals.
This cell line should be handled under Biosafety Level 2 (BSL-2) conditions based on the following characteristics:
- Human-derived origin — all human-derived cell lines require BSL-2 containment under Universal Precautions regardless of donor screening status; derived from a multiple myeloma patient
- Lentiviral transduction history — generated using lentiviral vectors; while replication-incompetent, standard BSL-2 containment for lentivirally transduced lines applies until confirmed negative for replication-competent lentivirus (RCL)
- Bloodborne pathogen risk — human-derived material of hematopoietic origin; treat as potentially infectious for EBV, Hepatitis, and other bloodborne pathogens
Users must consult their institutional biosafety committee (IBC) prior to use. Individual institutions may impose additional requirements based on local policy.
- Human-derived hematopoietic material — treat as potentially infectious; follow BSL-2 guidelines
- Lentiviral transduction history — handle in accordance with IBC guidelines for lentivirally transduced suspension cell lines
- Suspension growth — aerosol risk is elevated with suspension cultures; use appropriate containment when pipetting or centrifuging
- DMSO (cryoprotectant) — skin-penetrating solvent present in freezing medium; wear nitrile gloves at all times during thawing
- Disposal — decontaminate all cell culture waste with 10% bleach prior to disposal in accordance with local biohazardous waste regulations
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