How should I thaw PromoCell normal human cells to obtain high viability?
To achieve high post‑thaw viability, please follow the protocol below for anchorage‑dependent primary human cells.
General thawing procedure
1. Remove the cryovial from liquid nitrogen and transport it to the cell culture laboratory on dry ice.
2. Thaw the vial in a 37°C water bath for approximately 2 minutes, until the contents are just defrosted. Keep the vial immersed up to, but not above, the screw cap.
3. Under a laminar flow hood, disinfect the outside of the vial thoroughly with 70% ethanol.
4. Transfer the thawed cell suspension into 9 ml of pre‑warmed culture medium (1:10 dilution).
Seeding options: Proceed with one of the following options:
Option A: Direct plating
For cell‑type‑specific handling recommendations, please refer to the Instruction Manual of the respective cell type, as thawing and seeding requirements may vary.
General thawing procedure
1. Remove the cryovial from liquid nitrogen and transport it to the cell culture laboratory on dry ice.
2. Thaw the vial in a 37°C water bath for approximately 2 minutes, until the contents are just defrosted. Keep the vial immersed up to, but not above, the screw cap.
3. Under a laminar flow hood, disinfect the outside of the vial thoroughly with 70% ethanol.
4. Transfer the thawed cell suspension into 9 ml of pre‑warmed culture medium (1:10 dilution).
Seeding options: Proceed with one of the following options:
Option A: Direct plating
- Plate the diluted cell suspension directly at the recommended seeding density.
- Change the medium after 16–24 hours.
- Centrifuge the diluted cell suspension, discard the supernatant, and resuspend the cell pellet in 1 ml of fresh medium.
- Plate the cells at the recommended seeding density.
- Change the medium no earlier than 24 hours after seeding.
For cell‑type‑specific handling recommendations, please refer to the Instruction Manual of the respective cell type, as thawing and seeding requirements may vary.
Products relevant to this topic
Normal Human Epidermal Keratinocytes (NHEK) juvenile foreskin, single donor
Cat No
C-12001
Primary Human Keratinocytes isolated from the epidermis of juvenile foreskin or adult skin from single or pooled donors.
Normal Human Epidermal Keratinocytes (NHEK) adult, single donor
Cat No
C-12003
Primary Human Keratinocytes isolated from the epidermis of juvenile foreskin or adult skin from single or pooled donors.
Normal Human Epidermal Keratinocytes (NHEK) juvenile foreskin, pooled
Cat No
C-12005
Primary Human Keratinocytes isolated from the epidermis of juvenile foreskin or adult skin from single or pooled donors.
Normal Human Epidermal Keratinocytes (NHEK) adult, pooled
Cat No
C-12006
Primary Human Keratinocytes isolated from the epidermis of juvenile foreskin or adult skin from single or pooled donors.
Normal Human Epidermal Keratinocytes (NHEK), juvenile foreskin isolated in Growth Medium 3, single donor
Cat No
C-12011
Primary Human Keratinocytes isolated in serum-free and BPE-free Keratinocyte Growth Medium 3. Available from juvenile foreskin or adult skin from single or pooled donors.
Normal Human Epidermal Keratinocytes (NHEK) adult, isolated in Growth Medium 3, single donor
Cat No
C-12013
Primary Human Keratinocytes isolated in serum-free and BPE-free Keratinocyte Growth Medium 3. Available from juvenile foreskin or adult skin from single or pooled donors.
Normal Human Epidermal Keratinocytes (NHEK) juvenile foreskin, isolated in Growth Medium 3, pooled
Cat No
C-12015
Primary Human Keratinocytes isolated in serum-free and BPE-free Keratinocyte Growth Medium 3. Available from juvenile foreskin or adult skin from single or pooled donors.
Normal Human Epidermal Keratinocytes (NHEK) adult, isolated in Growth Medium 3, pooled
Cat No
C-12016
Primary Human Keratinocytes isolated in serum-free and BPE-free Keratinocyte Growth Medium 3. Available from juvenile foreskin or adult skin from single or pooled donors.