Human Normal Peripheral Blood Mononuclear Cells line ATCC® PCS800011
What media does recommend for the culture of ATCC® PCS-800-011 ™, Human Peripheral Blood Mononuclear (PBMC) and how to subculture and Maintenance PBMC
I have Cryopreserved Human Peripheral Blood Mononuclear Cells from atccc and only that protocol mention about thawing media (Hank's Balanced Salt Solution without Ca2+ orMg2+ supplemented with 10% Fetal Bovine Serum )
But which media to use for culture
How long are the cells viable after Post-thaw Cryopreserved HPBMC ? And can I store them in refrigerator for further use in the next day
Becouse iam having only one vial of Cryopreserved HPBMC and I want to use it to determine the effect of selected samples on cytokine production using the respective enzyme immunoassay kits (ELISA)
Here is Q&A from ATCC about culturing PBMC:
Culture medium for PCS-800-011 and PCS-800-013
What media does ATCC recommend for the culture of ATCC® PCS-800-011 ™, Human Peripheral Blood Mononuclear (PBMC) and ATCC® PCS-800-013 ™, Human Bone Marrow Mononuclear cells?
ATCC does not recommend maintaining mononuclear cells in culture in the absence of application-specific growth factors. We have found that mononuclear cells can however, be maintained in un-supplemented RMPI 1640 (ATCC® 30-2001 ™) for several days with high viability, although the immunophenotype will be altered relative to that immediately post-thaw. Adding 10% heat inactivated serum in RPMI may help.
An article is attached for more info.
- Generally speaking, dealing with PBMC cultures requires special media that can sustain it viability and maintain their growth potentiality. Originating from myeloid and lymphoid stem cells basement, it was found among other media formulae, that RPMI 1640 is one of the best nutritive culture media used. Complete RPMI 1640 that contains sodium bicarbonate, L-glutamate and serum is the best to use. However, Fully supplemented RMPI 1640 medium without serum are routinely used after addition of 10% FCS or BCS.
- Based on the quality of the preservation medium, technicality of the preservation and post-thawing processing of PBMC and other cell lines, the recovery and the viability percentage varies.
- Though, immediate culturing of the thawed cells is recommended, leaving of suspensions of the PBMC cell lines overnight at 2-8 degree Celsius is possible considering possible loss of their viability. Continuous parallel subculturing of the cells with preservation into high concentration-small volume quantities (6-12 X 1000000/mL) while performing normal would be the best way to guarantee RBMC sourcing.
- It was found that many cell lines preservation formulae could be used for storage purposes including: 70%RBMI 1640 + 20% FCS/BCS + 10% DMSO; 80% RPMI 1640 + 20% FCS/BCS; 80% serum devoid RPMI 1640 + 20% glycerol.
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