Catalog Number B1016
Product Name Neonatal rat dermal fibroblasts
Storage Liquid Nitrogen
Product Format Frozen vial
Cells Number >90% confluent in Frozen Vial
*Caution: Although primary cells are tested pathogen-free, investigators should handle these cells with caution and treat all animal cells as
potential pathogens, since no test procedure can completely guarantee the absence of infectious agents. Proper precautions must be taken to
avoid exposure. Always wear proper protective equipment (Gloves, safety glasses, etc.) when handling these materials. We recommend
following the universal procedures for handling products of animal origin as the minimum precaution against contamination.
Neonatal rat dermal fibroblasts cells were isolated from normal neonatal SD rat skin tissue. The cells are
shipped in proliferating culture or frozen vial with a confluence of > 90% (the cells are provided at
passage 1). ENDO-Growth medium (EGM-2102) containing 5% serum and growth supplement is
recommended for cell culture and these cells have an average minimum population doubling levels > 8
when cultured following the detailed protocol. Cells are tested negative for common experimental
animal pathogens, and mycoplasma in vitro. When you receive the cells, leave the flask in 37°C C02
incubator for 1 hour first and then replace the transport medium with fresh ENDO-Growth medium
(EGM-2102). Let the cells grow for 24 hours before subculture.
PECAM1 >95% positive by immunofluorescence
VE-Cadherin >95% positive by immunofluorescence
Neonatal rat dermal fibroblasts Negative for mycoplasma
PRODUCT USE AND SHIPPING STATUS
Product Use Neonatal rat dermal fibroblasts cells are for research only
Shipping Status Frozen vial
1) Coating T25 flasks. Add 2 ml AlphaBioCoat (AC001) into a T25 flask and ensure entire interior
surface is coated with the solution. After 30 minutes, dispose of AlphaBioCoat (AC001) by
aspiration. Gently rinse and aspirate flask with Phosphate Buffer Solution (1XPBS-001). The flask
is now ready for use (no need for overnight incubation when coated with AC001)
2) If you are using the coated flask the same day, add about 4 ml of Endo-Growth media to the
coated flask. If the media changes color from pink to yellow, aspirate and discard the media.
Add 4ml of fresh media to the coated flask.
3) Thaw the cells in a 37°C water bath. Once you see a small amount of ice left in the vial, spray the
vial with 70% Ethanol and wipe it down.
4) Transfer the vial into your Biosafety cabinet.
5) Using a 2 or 5ml pipet, pipet the cells out of the vial.
6) Transfer your cell suspension in to your coated plate that have the 4 ml media in it.
7) You should have a total working volume of 5ml of cell suspension in the flask; close the cap.
Make sure cells are evenly distributed in the flask by moving the flask left and right five times.
Move it up and down for and additional five times.
8) Place flask in a 37°C incubator with 5% C02. If flask is not vented, please loosen cap.
9) Change media after 48 hours.