WTF, MCF10A??? how can you sit in trypsin for 30 bloody minutes and *still* not detach? honestly you are THE WORST. i thought MDCK were bad enough, well little did i know. well come to think of it, maybe they *were* the worst, because they would also require trypsin for about half an hour but they would look a bit damaged the next day, so i figured out my own protocol in the end. MCF10A though, i’m impressed, i’m very fucking impressed. literally 10 minutes after i add medium, they drop down, attach and begin spreading. no, you cannot pipette them up, they’re properly attached and all. basically the cell suspension that you needed for the experiment, is gone. WTF???
except *of course* when i need them to attach and spread, like i’ve been waiting for um, 7 hours, and they’re still kinda rounded? which is not what i need? oh okay, i get it, you don’t like bare greiner’s plastic (honestly i don’t blame you, i’m not particularly fond of it myself, but it’s all we’ve got so please work with me here? MCF10A? please?). but you can’t seriously think that will stop a cell biologist with a shitton of experience, right? you want matrix? i’ll give you matrix, i’ll seed you @300K, @500K if needs be, and then i will replate you into the same wells @50K for the experiment, ah? howaboutthat? oh and i’ll coat a couple of wells with fibronectin as well, just to cover all the bases. pfffft.
okay i need to change my workstuff tag to “angrycellbiology”. yes it’s a rip-off but idc.Â
oh hey, joan brugge also recommends trypsin for 30 min, heh. so i guess there is no way around it then. UGH.
ETA: fucci is go, fingers crossed.
