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KCL-ENG-101
⚠ FOR RESEARCH USE ONLY — Not for diagnostic or therapeutic procedures
KCL-ENG-101 is a HeLa cell line (human cervical adenocarcinoma, epithelial, adherent) characterized and validated for endogenous expression of human Angiotensin-Converting Enzyme 2 (ACE2). Unlike the RFP-tagged ACE2 lines in this series, KCL-ENG-101 expresses ACE2 from its native genomic locus without transgenic modification or antibiotic selection — making it the most physiologically representative model for endogenous ACE2 receptor biology.
ACE2 is confirmed at moderate endogenous expression levels: 92% of cells are ACE2-positive by flow cytometry (MFI greater than 2-fold over isotype control), ACE2 mRNA is detected at Ct 26 to 28 by qPCR, and Western blot shows bands at approximately 120 kDa (glycosylated) and 92 kDa (deglycosylated). The line is suitable for SARS-CoV-2 pseudovirus entry assays, ACE2 inhibition screening, ACE2 receptor biology studies, and co-culture models. All lots are quality tested prior to release for ACE2 expression, viability, mycoplasma, and sterility.
Note: HeLa cells do not natively express TMPRSS2. For assays requiring Spike protein priming via TMPRSS2-dependent entry, co-express TMPRSS2 by transient transfection or supplement with exogenous trypsin.
DMEM supplemented with 10% heat-inactivated FBS and 1% Penicillin/Streptomycin. No antibiotic selection is required — this line expresses ACE2 endogenously and does not carry a selection marker. Subculture at 80 to 90% confluence using 0.25% Trypsin-EDTA; split 1:3 to 1:6 every 2 to 3 days at 37°C, 5% CO₂.
All lots are quality tested prior to release. ACE2 expression is confirmed at the protein, mRNA, and surface level for each production lot.
| Parameter | Result | Specification |
|---|---|---|
| ACE2 surface expression (flow cytometry) | ✓ Positive | 92% positive; MFI >2× isotype control (anti-ACE2, clone 171606) |
| ACE2 mRNA (qRT-PCR) | ✓ Detected | Ct 26 to 28 (endogenous); GAPDH Ct 18 to 20 as reference |
| ACE2 protein (Western blot) | ✓ Confirmed | Band at ~120 kDa (glycosylated) and ~92 kDa (deglycosylated); anti-ACE2 antibody |
| Post-thaw viability | ✓ Pass | ≥85% by Trypan Blue exclusion |
| Passage number at freeze | ✓ Confirmed | P4 to P6 from original thaw |
| Marker | Result | Notes |
|---|---|---|
| Cytokeratin 18 (CK18) | ✓ Positive | Epithelial lineage marker; confirms HeLa identity |
| CD13 (ANPEP) | ✓ Positive | Alternative coronavirus receptor; present in HeLa |
| HLA Class I | ✓ Positive | Immune-competent background; useful for T-cell co-culture assays |
| TMPRSS2 | ≈ Low / Negative | Not natively expressed in HeLa; co-express or add exogenous trypsin for Spike-primed entry |
| CD45 | ✗ Negative | Confirms absence of hematopoietic contamination |
| Test | Result | Method |
|---|---|---|
| Mycoplasma | ✗ Negative | PCR and luminescence assay |
| Sterility — Bacteria & Fungi | ✗ Negative | Culture; no bacterial or fungal growth |
Certificates of Analysis with lot-specific QC data including flow cytometry histograms and qPCR data are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-101 is shipped as a cryopreserved vial on dry ice. These are adherent HeLa cells requiring no antibiotic selection. Follow the steps below upon receipt.
Subculture at 80 to 90% confluence. Remove medium, rinse with PBS, add 0.25% Trypsin-EDTA and incubate 2 to 3 minutes at 37°C. Neutralize with complete medium and split 1:3 to 1:6 every 2 to 3 days. No selection antibiotic is required. Verify ACE2 expression periodically by flow cytometry if extended culture is planned. Full protocols are available on the Kulture Scientific protocol page.
KCL-ENG-101 is available as a cryopreserved vial containing at least 5×10&sup5; viable cells in cryopreservation medium (90% FBS + 10% DMSO).
| Product | Cat. No. | Format | Cell Number |
|---|---|---|---|
| Human ACE2 Expression in HeLa Cell Line | KCL-ENG-101 | Cryopreserved Vial | ≥5×105 cells/vial |
- Storage — store in liquid nitrogen vapor phase (−150°C to −196°C); cells are stable for more than 5 years when cryopreserved correctly
- 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 ACE2 expression; thaw only the quantity needed for immediate use
- Shipping condition — cryopreserved vials are shipped on dry ice; inspect packaging upon receipt and transfer to liquid nitrogen storage immediately
- Cryopreservation medium — cells are cryopreserved in 90% FBS + 10% DMSO; wear nitrile gloves during thaw procedures and avoid skin contact with DMSO
- No selection antibiotic required — this is an endogenous expression line; do not add Puromycin or other selection antibiotics during routine culture
- Certificate of Analysis — lot-specific QC documentation including ACE2 flow cytometry data, qPCR results, viability, and sterility 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 — HeLa cells are derived from human cervical adenocarcinoma tissue; all human-derived cell lines require BSL-2 containment under Universal Precautions
- HPV-18 sequences — HeLa cells are known to harbor integrated HPV-18 sequences; handle accordingly with BSL-2 precautions at all times
- ACE2 expression — this line expresses the SARS-CoV-2 entry receptor at endogenous levels; if used with viral constructs (e.g., VSV-G pseudovirus) or live SARS-CoV-2, follow institutional BSL-2 or BSL-3 guidelines as appropriate for the viral component
Users must consult their institutional biosafety committee (IBC) prior to use, particularly when this line is used in combination with pseudoviral or live-virus systems.
- Human-derived material with HPV-18 — treat as potentially infectious; work exclusively in a certified biosafety cabinet; follow BSL-2 guidelines
- ACE2 receptor expression — endogenous ACE2 makes these cells susceptible to SARS-CoV-2 Spike-mediated entry in pseudovirus assays; ensure appropriate containment for any viral component used
- DMSO (cryoprotectant) — present in cryopreservation medium; skin-penetrating solvent; wear nitrile gloves during thawing
- Waste 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-101 is a HeLa cell line (human cervical adenocarcinoma, epithelial, adherent) characterized and validated for endogenous expression of human Angiotensin-Converting Enzyme 2 (ACE2). Unlike the RFP-tagged ACE2 lines in this series, KCL-ENG-101 expresses ACE2 from its native genomic locus without transgenic modification or antibiotic selection — making it the most physiologically representative model for endogenous ACE2 receptor biology.
ACE2 is confirmed at moderate endogenous expression levels: 92% of cells are ACE2-positive by flow cytometry (MFI greater than 2-fold over isotype control), ACE2 mRNA is detected at Ct 26 to 28 by qPCR, and Western blot shows bands at approximately 120 kDa (glycosylated) and 92 kDa (deglycosylated). The line is suitable for SARS-CoV-2 pseudovirus entry assays, ACE2 inhibition screening, ACE2 receptor biology studies, and co-culture models. All lots are quality tested prior to release for ACE2 expression, viability, mycoplasma, and sterility.
Note: HeLa cells do not natively express TMPRSS2. For assays requiring Spike protein priming via TMPRSS2-dependent entry, co-express TMPRSS2 by transient transfection or supplement with exogenous trypsin.
DMEM supplemented with 10% heat-inactivated FBS and 1% Penicillin/Streptomycin. No antibiotic selection is required — this line expresses ACE2 endogenously and does not carry a selection marker. Subculture at 80 to 90% confluence using 0.25% Trypsin-EDTA; split 1:3 to 1:6 every 2 to 3 days at 37°C, 5% CO₂.
All lots are quality tested prior to release. ACE2 expression is confirmed at the protein, mRNA, and surface level for each production lot.
| Parameter | Result | Specification |
|---|---|---|
| ACE2 surface expression (flow cytometry) | ✓ Positive | 92% positive; MFI >2× isotype control (anti-ACE2, clone 171606) |
| ACE2 mRNA (qRT-PCR) | ✓ Detected | Ct 26 to 28 (endogenous); GAPDH Ct 18 to 20 as reference |
| ACE2 protein (Western blot) | ✓ Confirmed | Band at ~120 kDa (glycosylated) and ~92 kDa (deglycosylated); anti-ACE2 antibody |
| Post-thaw viability | ✓ Pass | ≥85% by Trypan Blue exclusion |
| Passage number at freeze | ✓ Confirmed | P4 to P6 from original thaw |
| Marker | Result | Notes |
|---|---|---|
| Cytokeratin 18 (CK18) | ✓ Positive | Epithelial lineage marker; confirms HeLa identity |
| CD13 (ANPEP) | ✓ Positive | Alternative coronavirus receptor; present in HeLa |
| HLA Class I | ✓ Positive | Immune-competent background; useful for T-cell co-culture assays |
| TMPRSS2 | ≈ Low / Negative | Not natively expressed in HeLa; co-express or add exogenous trypsin for Spike-primed entry |
| CD45 | ✗ Negative | Confirms absence of hematopoietic contamination |
| Test | Result | Method |
|---|---|---|
| Mycoplasma | ✗ Negative | PCR and luminescence assay |
| Sterility — Bacteria & Fungi | ✗ Negative | Culture; no bacterial or fungal growth |
Certificates of Analysis with lot-specific QC data including flow cytometry histograms and qPCR data are available upon request. Contact Kulture Scientific with your lot number.
KCL-ENG-101 is shipped as a cryopreserved vial on dry ice. These are adherent HeLa cells requiring no antibiotic selection. Follow the steps below upon receipt.
Subculture at 80 to 90% confluence. Remove medium, rinse with PBS, add 0.25% Trypsin-EDTA and incubate 2 to 3 minutes at 37°C. Neutralize with complete medium and split 1:3 to 1:6 every 2 to 3 days. No selection antibiotic is required. Verify ACE2 expression periodically by flow cytometry if extended culture is planned. Full protocols are available on the Kulture Scientific protocol page.
KCL-ENG-101 is available as a cryopreserved vial containing at least 5×10&sup5; viable cells in cryopreservation medium (90% FBS + 10% DMSO).
| Product | Cat. No. | Format | Cell Number |
|---|---|---|---|
| Human ACE2 Expression in HeLa Cell Line | KCL-ENG-101 | Cryopreserved Vial | ≥5×105 cells/vial |
- Storage — store in liquid nitrogen vapor phase (−150°C to −196°C); cells are stable for more than 5 years when cryopreserved correctly
- 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 ACE2 expression; thaw only the quantity needed for immediate use
- Shipping condition — cryopreserved vials are shipped on dry ice; inspect packaging upon receipt and transfer to liquid nitrogen storage immediately
- Cryopreservation medium — cells are cryopreserved in 90% FBS + 10% DMSO; wear nitrile gloves during thaw procedures and avoid skin contact with DMSO
- No selection antibiotic required — this is an endogenous expression line; do not add Puromycin or other selection antibiotics during routine culture
- Certificate of Analysis — lot-specific QC documentation including ACE2 flow cytometry data, qPCR results, viability, and sterility 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 — HeLa cells are derived from human cervical adenocarcinoma tissue; all human-derived cell lines require BSL-2 containment under Universal Precautions
- HPV-18 sequences — HeLa cells are known to harbor integrated HPV-18 sequences; handle accordingly with BSL-2 precautions at all times
- ACE2 expression — this line expresses the SARS-CoV-2 entry receptor at endogenous levels; if used with viral constructs (e.g., VSV-G pseudovirus) or live SARS-CoV-2, follow institutional BSL-2 or BSL-3 guidelines as appropriate for the viral component
Users must consult their institutional biosafety committee (IBC) prior to use, particularly when this line is used in combination with pseudoviral or live-virus systems.
- Human-derived material with HPV-18 — treat as potentially infectious; work exclusively in a certified biosafety cabinet; follow BSL-2 guidelines
- ACE2 receptor expression — endogenous ACE2 makes these cells susceptible to SARS-CoV-2 Spike-mediated entry in pseudovirus assays; ensure appropriate containment for any viral component used
- DMSO (cryoprotectant) — present in cryopreservation medium; skin-penetrating solvent; wear nitrile gloves during thawing
- Waste disposal — decontaminate all cell culture waste with 10% bleach prior to disposal in accordance with local biohazardous waste regulations
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