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Found 4 matching solutions for this experiment
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Normal human epidermal keratinocytes were isolated from neonatal skin. Frozen keratinocytes were thawed and cultivated at clonal density on a feeder layer of irradiated or mitomycin C–treated 3T3-J2 cells, at 37°C and 10% CO2 in a 3:1 mixture of the DMEM and Ham’s F12 medium supplemented with 10% FCS, 5 µg/ml insulin, 0.4 µg/ml hydrocortisone, 10−10 M cholera toxin, and 2 × 10−9 M triiodothyronine. Human epidermal keratinocytes were also cultivated with CnT-PR medium (CELLnTEC), EpiLife medium containing supplement S7 (Life Technologies), and MCDB153 medium containing bovine pituitary extract (Hashimoto et al., 1994). Keratinocytes were used between passages 2 and 7. The medium was changed every 4 d. |
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| Protocol tips |
| Normal human epidermal keratinocytes were isolated from neonatal skin. Frozen keratinocytes were thawed and cultivated at clonal density on a feeder layer of irradiated or mitomycin C–treated 3T3-J2 cells, at 37°C and 10% CO2 in a 3:1 mixture of the DMEM and Ham’s F12 medium supplemented with 10% FCS, 5 µg/ml insulin, 0.4 µg/ml hydrocortisone, 10−10 M cholera toxin, and 2 × 10−9 M triiodothyronine. Human epidermal keratinocytes were also cultivated with CnT-PR medium (CELLnTEC), EpiLife medium containing supplement S7 (Life Technologies), and MCDB153 medium containing bovine pituitary extract (Hashimoto et al., 1994). Keratinocytes were used between passages 2 and 7. The medium was changed every 4 d. |
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Third passage Neonatal Human Epidermal Keratinocytes (NHEKs Lonza) were initially cultured in serum-free conditions in Lonza's KGM-Gold media (basal medium supplemented with bovine pituitary extract, human epidermal growth factor, bovine insulin, hydrocortisone, Gentamicin, Amphotericin-B, Epinephrine and Transferrin) in collagen coated (1∶100; Type 1 rat tail collagen (Sigma) : PBS) T25 flasks. The cells were cultured in a 37°C incubator with 5% CO2, with the media being changed every 2 days until approximately 80% confluent. |
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| Protocol tips |
| Third passage Neonatal Human Epidermal Keratinocytes (NHEKs Lonza) were initially cultured in serum-free conditions in Lonza's KGM-Gold media (basal medium supplemented with bovine pituitary extract, human epidermal growth factor, bovine insulin, hydrocortisone, Gentamicin, Amphotericin-B, Epinephrine and Transferrin) in collagen coated (1∶100; Type 1 rat tail collagen (Sigma) : PBS) T25 flasks. The cells were cultured in a 37°C incubator with 5% CO2, with the media being changed every 2 days until approximately 80% confluent. |
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Protocol tips |
Downstream tips |
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Frozen keratinocytes were thawed and cultivated at clonal density on a feeder layer of irradiated or mitomycin C–treated 3T3-J2 cells, at 37°C and 10% CO2 in a 3:1 mixture of the DMEM and Ham’s F12 medium supplemented with 10% FCS, 5 µg/ml insulin, 0.4 µg/ml hydrocortisone, 10−10 M cholera toxin, and 2 × 10−9 M triiodothyronine |
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| Protocol tips |
| Frozen keratinocytes were thawed and cultivated at clonal density on a feeder layer of irradiated or mitomycin C–treated 3T3-J2 cells, at 37°C and 10% CO2 in a 3:1 mixture of the DMEM and Ham’s F12 medium supplemented with 10% FCS, 5 µg/ml insulin, 0.4 µg/ml hydrocortisone, 10−10 M cholera toxin, and 2 × 10−9 M triiodothyronine |
| Upstream tips |
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High performance, scalability and quality, ensuring purity and safety of the final product.
Optimized for transient transfection and infection or stable expression in suspension
100% chemically defined, animal component-free, serum-free.
Manufactured under ISO 9001 & ISO 13485 quality standards |
Growth in gene therapies and viral vector vaccines has increased the demand for robust HEK293 cell lines. Sartorius offers four high-performing suspension HEK media that are designed for use with the high variability of HEK293 cell lines, the versatility of adeno-associated virus and the complexity of viral vector processes. The portfolio also includes a unique feed supplement to boost viral production for gene therapies and vaccines. |
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| Upstream tips |
High performance, scalability and quality, ensuring purity and safety of the final product.
Optimized for transient transfection and infection or stable expression in suspension
100% chemically defined, animal component-free, serum-free.
Manufactured under ISO 9001 & ISO 13485 quality standards |
| Protocol tips |
| Growth in gene therapies and viral vector vaccines has increased the demand for robust HEK293 cell lines. Sartorius offers four high-performing suspension HEK media that are designed for use with the high variability of HEK293 cell lines, the versatility of adeno-associated virus and the complexity of viral vector processes. The portfolio also includes a unique feed supplement to boost viral production for gene therapies and vaccines. |
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