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Cnt prime media

Manufactured by CELLnTEC
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CnT-Prime media is a cell culture medium designed for the growth and maintenance of a variety of cell types. It provides the necessary nutrients and components to support cell proliferation and viability in an in vitro environment.

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Lab products found in correlation

0.4 μm pore inserts, by Merck Group (4 mentions) Cnt 07 media, by CELLnTEC (4 mentions) Cellstart, by Thermo Fisher Scientific (1 mentions) Cnt pr 3d differentiation medium, by CELLnTEC (1 mentions) Cnt prime medium, by CELLnTEC (1 mentions) Epilife kg2, by Kurabo (1 mentions) Dmem f12 media, by Thermo Fisher Scientific (1 mentions)

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CnT-Prime (20 citations)

6 protocols using cnt prime media

1

Keratinocyte Culture for Epidermal Barrier Development

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Example 4

Keratinocytes are seeded at a density of 2.0-2.5×105 cells/cm2 of polyethylene terephthalate (PET) membrane with 0.4 μm pore inserts (EMD Millipore; Cat. No.: MCHT12H48) in CnT-07 media (CELLnTEC) or CnT-Prime media (CELLnTEC).

Day 3 (D3) after seeding, the media are switched to CnT-02-3D (CELLnTEC) or CnT-3D Barrier (CELLnTEC). On day 4, the cells air exposed by feeding the bottom of the insert with CnT-02-3D CnT-3D Barrier. From Day 4 onward, the epidermal layer is fed daily with CnT-02-3D or CnT-3D Barrier until harvested at Day 14.

US11739217B2. Engineered skin equivalent, method of manufacture thereof and products derived therefrom (2023-08-29). VitroLabs Inc [US], King's College London [GB]. Inventors: Ingvar Helgason [US], Dusko Ilic [GB].
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2

Keratinocyte Culture for Epidermal Layer Formation

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Example 4

Keratinocytes are seeded at a density of 2.0-2.5×105 cells/cm2 of polyethylene terephthalate (PET) membrane with 0.4 μm pore inserts (EMD Millipore; Cat. No.: MCHT12H48) in CnT-07 media (CELLnTEC) or CnT-Prime media (CELLnTEC).

Day 3 (D3) after seeding, the media are switched to CnT-02-3D (CELLnTEC) or CnT-3D Barrier (CELLnTEC). On day 4, the cells air exposed by feeding the bottom of the insert with CnT-02-3D CnT-3D Barrier. From Day 4 onward, the epidermal layer is fed daily with CnT-02-3D or CnT-3D Barrier until harvested at Day 14.

US10711136B2. Engineered skin equivalent, method of manufacture thereof and products derived therefrom (2020-07-14). VitroLabs Inc [US], King's College London [GB]. Inventors: Ingvar Helgason [US], Dusko Ilic [GB].
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3

Epidermal Keratinocyte Culture Protocol

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Example 4

Keratinocytes are seeded at a density of 2.0-2.5×105 cells/cm2 of polyethylene terephthalate (PET) membrane with 0.4 μm pore inserts (EMD Millipore; Cat. No.: MCHT12H48) in CnT-07 media (CELLnTEC) or CnT-Prime media (CELLnTEC).

Day 3 (D3) after seeding, the media are switched to CnT-02-3D (CELLnTEC) or CnT-3D Barrier (CELLnTEC). On day 4, the cells air exposed by feeding the bottom of the insert with CnT-02-3D CnT-3D Barrier. From Day 4 onward, the epidermal layer is fed daily with CnT-02-3D or CnT-3D Barrier until harvested at Day 14.

US11591471B2. Engineered skin equivalent, method of manufacture thereof and products derived therefrom (2023-02-28). VitroLabs Inc [US], King's College London [GB]. Inventors: Ingvar Helgason [US], Dusko Ilic [GB].
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4

Construction of Epidermal Equivalent Model

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The protocol for construction of the epidermal‐equivalent model was described previously.14, 15, 16 Normal human keratinocytes (NHEKs) from Kurabo (Osaka, Japan) were cultured in Epilife‐KG2 (Kurabo) containing 0.06 mM Ca2+. Millicells with hanging 0.4 μm PET inserts (Millipore, Billerica, MA) were incubated with CellStart (Invitrogen Life Technologies, Carlsbad, CA) in 50X DPBS dilution. Then, 2.2 × 105 keratinocytes/500 μL in CnT Prime media (CELLnTEC, Bern, Switzerland) were seeded and CnT Prime medium was added outside the inserts. After 3 days, the medium was removed and CnT‐PR‐3D differentiation medium (CELLnTEC) containing 50 μg/mL ascorbic acid was added both inside and outside the inserts. On the next day, cultures were lifted to the air‐medium interface by removing the medium inside the inserts and reducing the amount outside the inserts to 500 μL. Cultures were fed every day with 500 μL of differentiation medium.
Umino Y, & Denda M. (2023). Effect of red light on epidermal proliferation and mitochondrial activity. Skin Research and Technology, 29(9), e13447.
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5

Establishment of 3D Epidermal Barrier Model

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Example 4

Keratinocytes are seeded at a density of 2.0-2.5×105 cells/cm2 of polyethylene terephthalate (PET) membrane with 0.4 μm pore inserts (EMD Millipore; Cat. No.: MCHT12H48) in CnT-07 media (CELLnTEC) or CnT-Prime media (CELLnTEC).

Day 3 (D3) after seeding, the media are switched to CnT-02-3D (CELLnTEC) or CnT-3D Barrier (CELLnTEC). On day 4, the cells air exposed by feeding the bottom of the insert with CnT-02-3D CnT-3D Barrier. From Day 4 onward, the epidermal layer is fed daily with CnT-02-3D or CnT-3D Barrier until harvested at Day 14.

US11091639B2. Engineered skin equivalent, method of manufacture thereof and products derived therefrom (2021-08-17). VitroLabs Inc [US], King's College London [GB]. Inventors: Ingvar Helgason [US], Dusko Ilic [GB].
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6

Proliferation Assays for p63 Mutants

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For in vitro proliferation assay, freshly isolated p63+/+ and p63+/− PIMECs were plated into 24-well plates at 30,000–120,000 cells per well in DMEM/F12 media (Gibco), switched to CNT Prime media (Cellntec) after two days, and stained for the proliferation marker Ki67 either after 6 days (when 70,000–120,000 were plated) or after 12 days (when 30,000 cells were plated), as described below. Five random non-overlapping fields were photographed at ×20 magnification, and the percent of Ki67-positive cells was calculated. For the in vivo proliferation assay, mammary gland #4 from p63+/+ and p63+/− sisters were flattened on filter paper, fixed in formalin, embedded in paraffin, and sectioned (5 μm), followed by immunofluorescent staining as described below. Ten random non-overlapping fields were photographed at ×20 magnification, and the percent of Ki67-positive cells within YFP-positive cells was calculated.
Eyermann C.E., Li J, & Alexandrova E.M. (2021). p63 suppresses the ability of pregnancy-identified mammary epithelial cells (PIMECs) to drive HER2-positive breast cancer. Cell Death & Disease, 12(6), 525.
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