Mammalian cell culture media HEK

Cells are sourced from various tissues to grow them in in-vitro conditions. Therefore, cell specific nutrients are important for their survival, maintenance and growth. Determining the appropriate cell culture media is a challenge if you are growing a cell line or a microorganism for the first time. Established cell lines, primary cells, stem cells, bacteria and Yeast all require varied nutrients from basic to complex. Based on the cell type, one can easy find what media and nutrients your peers have used before you try to reinvent the wheel.

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CnT-Prime, Epithelial Culture Medium

Cellntec

Upstream tips	Protocol tips	Downstream 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.

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.

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.

Manufacturer protocol Publication protocol 1 Relevant paper

KGMTM Gold Keratinocyte Growth Medium BulletKitTM

Lonza

Upstream tips	Protocol tips	Downstream 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.

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.

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.

Manufacturer protocol Publication protocol 1 Relevant paper

EpiLife™ Medium, with 60 µM calcium

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream 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

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

Manufacturer protocol Publication protocol 1 Relevant paper

HEK GM

SARTORIUS

Upstream tips	Protocol tips	Downstream 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	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.

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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