Freezing and Thawing S2 Cells - Revision history

Reddj: added SOP requirements-JR

2020-06-05T14:28:18Z

added SOP requirements-JR

← Older revision		Revision as of 14:28, 5 June 2020
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			==SOP==
			*[[SOP- Cryogenic Materials]]
			*[[SOP- Biosafety Cabinets]]

			==METHOD==
	#Use and early passage (1-10) of exponentially growing (about ~10^7 cells).		#Use and early passage (1-10) of exponentially growing (about ~10^7 cells).
	#Detach by pipeting a stream of media over the cells		#Detach by pipeting a stream of media over the cells

Davebrid at 16:07, 31 July 2009

2009-07-31T16:07:22Z

← Older revision		Revision as of 16:07, 31 July 2009
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	see PMID 18388942		see PMID 18388942

			[[Category: Cell Culture]]

Davebrid: initial page

2009-07-31T16:07:08Z

initial page

New page

#Use and early passage (1-10) of exponentially growing (about ~10^7 cells).
#Detach by pipeting a stream of media over the cells
#Transfer into a 15 mL falcon tube
#Spin at 500g for 10 min, save the supernatant
#Prepare freezing media:
##Add 4.5 mL supernatant
##Add 4.5 mL fresh media
##Add 100 uL sterile DMSO
#Resuspend to 1/10th original culture volume
#Dispense 0.5 mL into sterile cryovials
#Place in isopropanol freezing chamber and store at -80 for about a day
#Transfer to liquid nitrogen
#Cells should be viable for >5 years

see PMID 18388942