I am recently working with human mammary epithelial cells(HMEC) which is new to me. I just wanted to acquire the cryo-preservation method of HMEC as they don't need FBS for there culturing. So for preservation what supplements should be used apart form HMEC growth media.
Dipti Debnath
Hi Nabiha,
Here is my answer.
HMEC Cryo-Preservation
HMEC cells should be frozen in Glycerol I freezing media. This is a combination of MCDB 170 culture media with 10% glycerol and 15% FCS. Cells should be maintained at a density of 106 cells/mL. After slowly freezing the cells overnight, store them in the vial in the vapor phase of liquid nitrogen.
Tips
- To achieve optimal cell densities, be persistent in changing culture medium daily
- For maximum proliferating subcultures, subculture HMEC before 90% confluency is reached
- Damage to HMEC cells can occur during the trypsinization step; damage may result from excessive exposure to Trypsin or too harsh pipetting
- The number of passages achieved is individual specific and vary with the methods employed by investigators
- Freshly thawed aliquot: Thawed cells are fragile and should be thawed in less than 1 minute. Do not pellet the cells with centrifugation to remove freezing media. Rather, the DMSO or Glycerol will be diluted to a low enough concentration by adding the thawed aliquot directly to a flask containing media
https://www.hmec-cells.com/cell-culture-information/
https://www.atcc.org/products/pcs-600-010
Regards,
Dipti Debnath
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