Passaging Cells: Cell Culture Basics

www.thermofisher.com/global/e...
The handbook and videos provide an introduction to cell culture, with a focus on maintaining cell health throughout the processes of culturing, freezing, thawing and passaging cells. In this video, we focus on how to passage cells.

[@rayseodium]

Our Professor is using these videos to teach us the lab routines while labs are closed due to quarantine greetings from Germany

[@thermofisher]

Guten Tag! Grüß Gott! And props to your professor for maintaining an educational environment through this crisis.

[@achilleasp9061]

same in Greece

[@molecularkitchen]

True

[@nidesharai9792]

same here in the UK

[@rionassetnoc]

Same in US

[@unculturedmeat]

Thank you for uploading this playlist. This is my dream job. Hope to get there one day!

[@thermofisher]

If you can believe it, you can achieve it!

[@supb1437]

Hey, I do this every day at my work. It’s extremely rewarding! Good luck!

[@jeeva1996]

@@supb1437 what's your job title?

[@jenb4285]

@@jeeva1996 probably research tech/ associate/ technologist

[@priyankajaikumar]

Did you get the dream job?

[@manishathakur8435]

I have done the same procedure when I was a intern at Thermofisher Scientific, Banglore in 2019. This video reminded me my best days... Thermofisher is the best place to gain knowledge and learn technical skills. Hope to work there again.

[@thermofisher]

Glad to know that you liked interning with us, Manisha. We wish you good luck for the future.

[@lidamaryageorge8298]

Could you tell me how exactly did you get in there as an intern? .... I'm currently a student studying BSc microbiology right now.

[@fidanfazilova3542]

Hi. I have never done this before. in 2 month, I'll be in intern for 2 weeks in Japan. Seems like, I 'll be participate in this experiment as well, but I have concerns. Is it really hard for one who have never done it before? Am i gonna adjust the environment easily?

[@ramkrishnapangi7711]

😂

[@manishathakur8435]

​@@lidamaryageorge8298 I have joined a PG Diploma course Biotechnology skill enhancement program. After completing 6months of theory classes, our resumes were sent to multiple biotech companies. Hence, I was hired by Thermofisher.

[@jennanicolewise3156]

THANK YOU. This video was very helpful!

[@thermofisher]

We're so glad to hear that! Just doing our part to support and uplift the scientific community. Happy New Year!

[@ansarisumaiya4643]

Thanks for uploading this video. This video helps in our assignment. 👍😌

[@thermofisher]

Thank you for watching. Glad our video could help you in your assignment. Subscribe and get notified for more latest uploads.

[@chizpa305]

Awesome. It would be nice to have other procedures also.

[@thermofisher]

Thanks! In addition to Passaging Cells, we also have Aspetic Techniques, Cell Culture, Freezing and Thawing Cells. Was there a different procedure that you were hoping to see? Please let us know.

[@kamranshekh9404]

@@thermofisher Do you have videos for some basic methods for cell culture such as isolation of RNA/DNA, protein etc

[@songmanne]

Thank you very much for this video! I've recently started my training and feel quite nervous for it, so I can quickly forget some details. Your video'll be helpful for me.

[@thermofisher]

Hello songmanne, it's always great to hear positive feedback. Have a great day!

[@user-md4hg9ld7w]

Can you please tell me what microscope is being used in this video?

[@May04bwu]

You make it look so easy. Damn bubbles in the medium are killing me.

[@thermofisher]

These dang bubbles are killing me (And I) Must profess, I need to sleep (Need to sleep) When I think about this I lose my mind Stay still we'll be fiiiiinnnneeeee Prep my culture one more time

[@grinsunsharma567]

same here

[@Tomogeny]

@@thermofisher HAHA

[@vercas.1664]

I agreee

[@fareehashaikh]

@@thermofisher this made me expect britney lyrics every time i click on your replies here

[@codycigar6542]

I just applied for a position doing this and I hope I get it! Thank you so much for the detailed and concise instructions! One question: is the same pipette being used or are they different for each solution used?

[@oritsejemiyoejuetueyin3711]

Different pipettes are being used for the different solution (to avoid contaminating the solutions)

[@codycigar6542]

@@oritsejemiyoejuetueyin3711 thanks! I figured as much just wasn't sure if it was seen as wasteful or something.

[@molecularkitchen]

Good explanation.

[@thermofisher]

Thank you! Happy to be a part of scientific discourse!

[@jayk.6956]

Hi, are some of the flasks and biohazardous waste containers you guys are using made of plastic?

[@thermofisher]

Thank you for your question Jay. Could you please contact our technical support team so they can best assist you with this question? To reach them, please go to thermofisher.com/askaquestion. Please link them to the video in question. Thank you!

[@tobymann473]

Very helpful

[@irhamninasharudin208]

May i know the difference between trypsinization and passaging methods?

[@simranmehta5335]

Trypsinization is the is the process of cell dissociation using trypsin, an enzyme which breaks down proteins, to dissociate adherent cells from the vessel in which they are being cultured. Trypsinization is a part of passaging method. A method by which we pass the cells ''when a cell line reaches the point of growth in culture where it covers most of the bottom of the culture container, or about a 90% confluency, the cells must be resuspended, washed, used experimentally, frozen for later use, or re-seeded for further expansion in new culture containers''.For passaging the cells we need to dissociate from the surface by using trypsin

[@thermofisher]

Hi Irhmnhzqi. Thank you for the question. Trypsinization is cell dissociation using the enzyme trypsin. Passaging is the process of removing the medium and transferring the cells from a previous culture into fresh growth medium. For additional questions, please contact our technical support team at thermofisher.com/askaquestion. Thank you!

[@molecularkitchen]

Trypsinization is the enzymatic dissociation of cells from culture plate as well as between cells and passage means removing of dead cells and buffer and other secreted metabolites and provide new medium and culture space. Passage number is important in some cases e.g. primary cells.

[@deepthid8436]

What is the role of heat inactivated serum . What happens we use without heat inactivation.?

[@kianashah7029]

It was great

[@maneleip1892]

Hi I'M really new to the cell culture is this one of the techniques of cancer's cell culture?

[@drn6741]

Yes

[@hifikla]

Ну да, меня именно так и учили работать.

[@marroonet]

круто

[@esteladapper8892]

I wish I could find a lab in Toronto, or around it, that would let me work for free just to gain experience in cell culture.

[@JYOtiRaNJanMANgaRaj]

❤❤❤❤❤

[@efalusi4822]

What Cell that is,

[@messiah6418]

What are the order of the procedures?

[@thermofisher]

Thanks for watching. We have a step-by-step protocol on how to subculture adherent cells. The protocol has been linked below. www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/subculturing-adherent-cells.html For additional technical questions, please email [e-mail].

[@dareflamee]

1:35 Why is she allowing the cap to be placed on their BSC surface, isn't it best to not give any chances of contamination by holding the cap with her finger on the same hand she holds the t-25?

[@thermofisher]

Thanks for the question, will get back to you shortly.

[@gaslatvia3568]

Cause the wind that experiment table brought was from top to bottom, if placed with the inner face on the top, it would be contaminated by the wind.

[@dareflamee]

@@gaslatvia3568 My suggestion was to hold on to the cap on the hand that also holds the t-25 flask, and not place it on the surface of the cabinet at all.

[@dareflamee]

But its alright, I guess this video is just for demostration purposes

[@hugoavila9600]

While it is probably okay to place the cap on a clean (UV + IPA) surface, I like to be safer and avoid any kind of unnecessary exposure with a surface I can't guarantee to be free of contaminants. I do this by unscrewing the cap with one hand and holding in a finger curl (pointer finger) while the thumb and middle and ring fingers lift the flask for the right hand to come in with the pipette aid. The same hand can put the cap back on by transferring the grip to the pointer and thumb after being placed down. This way avoids contact between the cap and the working surface.

[@dylan302]

Use isolator instead of using biosafety

[@boyinalabcoatboyinalabcoat393]

You guys realize that the procedure is super simple and the problem lies in the calculations? Like there are 200+ videos saying the exact same things an none say anything about cell counting

[@thermofisher]

Thanks for watching. We'll get back to you with a response as soon as we can.

[@boyinalabcoatboyinalabcoat393]

@@thermofisher wow i did not expect any, I an very gratefull

[@moeemmanuel9395]

Arithmetic mean of your counted cells times 10.000 (Neubauer) times the dilution = cells per mL

[@deepakkumar-dr9lj]

I got into stem cell manufacturing and left that jobb......

[@koglesswarielanggoben8515]

this supposed to be aseptic technique but why is the person doing is no t wearing her hair net and closing her face?

[@DrexisEbon]

Because those are not necessary in all conditions of culture especially if using a properly ventilated BSC and media containing antimicrobials.

[@thermofisher]

Thanks for watching the video. The woman is working within a biological safety cabinet, which is an enclosed workspace. Class II BSCs are recommended for use in cell culture. If you look close enough, you can see the clear sash is pulled down in the front and covering the workers face and body allowing only her hands and arms to work inside the cabinet. To understand how the BSC works and protects the cell culture from the outside environment, the customer can reference this video, also by Thermo Fisher Scientific: kzbin.info/www/bejne/pXrYiISnr9xmaJY

[@molecularkitchen]

Good observation, one should take utmost precaution in the cell culture room.

[@edgartapia9005]

I was thinking the same thing lmao

[@edgartapia9005]

@@DrexisEbon it depends in the situation, if this was a manufacturing situation, there would be some red flags, like not having BSC sleeves and the lack of hair net.

[@weimullerjohann9118]

You can cut up many spaghetti and then multiply them unknown. Cheeky rip off nonsense.

[@faezor3326]

What about meat balls