Nicely made video, thanks! You might also consider Gibco's 'Tryple' which is said to be more gentle on cells. Also, I would view cells on the microscope prior to detaching adherent cells, to see if they were confluent or not. If not confluent, then no need to trypsinize yet. Well made video!
Is it right way to use nonsterile tissuepaper to sterilize equipment with IPA? & Can we keep trypsin under cell culture bottle up to finish of trypsinization in incubator,what are its drawbacks?
nice to see they use sterile pipettes instead of cannulas, not concerned about the lid of the culture being left off so long in a BSL-2 style hood (they are not bench tops where contamination is a higher risk) they just move hepa filtered air around (risk is low). However the risk to touch the inside of the flip top lid of the trypson holding cylinder & then contaminate the next users cell line is a HUGE concern. really need to think about using a screw cap for this part.
You need to consider cell density and percentage of trypsin. For example A549 cells would talks longer than 293 HEK, but 293HEK at a high seeding density could take longer than 5 minutes
This video shows many things you should avoid during cell culture
Thank you so much
Nicely made video, thanks! You might also consider Gibco's 'Tryple' which is said to be more gentle on cells. Also, I would view cells on the microscope prior to detaching adherent cells, to see if they were confluent or not. If not confluent, then no need to trypsinize yet. Well made video!
Is it right way to use nonsterile tissuepaper to sterilize equipment with IPA? & Can we keep trypsin under cell culture bottle up to finish of trypsinization in incubator,what are its drawbacks?
Are any animals killed during the extraction of Trypsin from their pancreas ? Or can it be done without killing the animal ?
Why did they leave the lid off the flask for so long....that's just asking for contamination to go right in there.
nice to see they use sterile pipettes instead of cannulas, not concerned about the lid of the culture being left off so long in a BSL-2 style hood (they are not bench tops where contamination is a higher risk) they just move hepa filtered air around (risk is low). However the risk to touch the inside of the flip top lid of the trypson holding cylinder & then contaminate the next users cell line is a HUGE concern. really need to think about using a screw cap for this part.
Make sure not to use wristbands as shown in this video that´s not proper sterile technique.
Use sleeved covers as well
thank you so much from Morocco for the vidéo. I have just a question do you think 5 minutes it's too long, maybe 1-2 min sholud be enough.
Depends on the cell line.
@@ariajohn37 Thanks Aria ✌
What cell line is being passaged? I ask because the five minute trypsinization is fairly long, so I wonder if the cells are a very adherent bunch.
You need to consider cell density and percentage of trypsin.
For example A549 cells would talks longer than 293 HEK, but 293HEK at a high seeding density could take longer than 5 minutes
The cells must not be left without medium or Trypsin solution for 5 mins
Left that T75 flask cap on the bench... suppose to hold it upside down Yikes !
Nooo, don´t spray into the LAF, this hurts the filters.
And the caps of the flask with the open side up!
look at that furry bracelet on the right wrist :)
pie pet
WAW!! If you perform cell culture for pharma purposes, don't look this video.Nevertheless, I "like" for the clownish effect... ;-)