Introduction to cell culture, splitting cells using trypsin and counting them using a hemocytometer
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- Опубліковано 3 лют 2025
- This video provides you with a general overview of the procedures typically used to "spit" a culture of immortalized adherent human cells maintained in tissue culture in a T75 flask.
This professor was my favorite professor and made the greatest impact on my life. I really wouldn't be the scientist I am today without his guidance and teaching.
Hello can you plss tell me application of immunohistochemistry
Regarding using the antibiotics for primary cells : Does is it necessary to add antibiotics whenever we change the media( let us say every 24 hours) for the first passage P0 , or it is recommended to add antibiotics just for the first 24 hours ?
@@funny11744 Antibiotic use in primary cultures is a balancing act. While they guard against contamination, especially in the crucial P0 stage, they can also affect cell behavior. To minimize these impacts, consider using antibiotics only for the first 24-48 hours of P0, then relying on strict aseptic technique for subsequent passages. Regularly monitor cultures for contamination and consult your protocol or a cell biologist for specific recommendations based on your cell type and research goals.
Thank you so much 🙏🙏🙏❤❤❤❤❤❤❤❤❤❤
Great video and explanations! Thank you for uploading it. I really enjoyed the camera technique, because is like you are doing it yourself.
TYSM for making this video. I'm currently applying for entry-level positions in biotech and this is one of the most sought-after techniques. This video is helping me identify the similarities between the cell culture I do now mammalian.
Very helpful video. Very well explained with examples and live demonstrations. Thank you❤
Very thorough tutorial !Thanks teacher !
This helped me alot! Thank you maam ! ❤ may you live a long and meaningful life ❤❤❤😊
braaaaaava you are the best one who was explanation cell culture to me >>> thanks a lot
Happy to hear that!
Thank u Olga 💓...keep posting, really helpful video 😊
I am so greatful for this helpful video! Thank you so much
Great video, thanks! Very helpul for my first times in the lab
best ever explanation
You are a hero to me, thank you.
Amazing video, Thank you for your effort. 👍👍👍
Thank you very much for this teaching..
Thank you so much for this good video. It make me clearly understand about cell culture. 😊❤🍀👩🔬
this video made my concept clear. thanks
You're welcome!
well prepared and clear
clear explanation indeed .
Thank you, it was amazing
Such a good video! Thank you :)
Nice video, my only recommendation is to wash the cells with dPBS (lacking Mg and Ca salts) rather than PBS prior to trypsin. Those divalent cations inhibit/slow down trypsin. I always wash at least two, sometimes three times, to speed up the trypsin step, which sometimes takes like 20min for my weird cells. Is It is a super cheap reagent, so not an issue to be wasteful.
Great point!
thanks olga soto
Very insightful . Thank u
What about renoving the neutralized trypsin and adding fresh media ?
Now that you have the number of cells per whatever volume (mL), how do you calculate the correct volume to seed a desired number of cells?
Tysm for the video❤
Super super thank you for sharing it with us… do you have experience with BREAST CANCER STEM CELLS???
More trypsin may be necessary depending on the potency of the trypsin as well
Cell detachment with trypsin depends on its potency - more active enzymes per volume means less solution is needed. To minimize cell damage, use the lowest amount of a high potency trypsin solution that effectively detaches your specific cell line.
At the minute 0:47 it is written ,, Immortalized cells could ....indefinetely" . Somebody could explain ?
It says "reproduce indefinitely." It means when these immortalized cells are grown under favorable condition they can reproduce indefinitely.
@@kosheeka thanks
Do you make other videos for MTT assay
THANK U SO MUCH
What is the name of that aspirator used in discarding ?
This is a lab vacuum aspirator. The tubes have filters attached to prevent any contamination that may occur.
Are any animals killed during the extraction of Trypsin from their pancreas ? Or can it be done without killing the animal ?
There are two main methods to produce the trypsin used in tissue culture. One is by purifying it from the pancreas of pigs; those pigs are killed by the thousands for human consumption, so in a way this uses a part of the animal usually considered of no value. The other method is by expressing it in recombinant expression systems. In this alternative method, no animal byproducts are used.
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Please mention how to count and distinguish ADSC ( Adipos derived stem cell) from other cells from SVF , using HEMOCYTOMETER and microscope. What total magnification of microscope îs required ? Thanks
Here's a quick tip:
• Use Trypan Blue to count viable cells in your hemocytometer (only unstained count!).
• Look for spindle-shaped cells under the microscope (100x-400x magnification). These are likely your ADSCs!
This separates them from round blood cells and cobblestone-shaped endothelial cells in SVF.
@@kosheeka many thanks
great job, I liked and sub.. it
helpful!
every time i see someone with bare skin (like the arm not covered by the lab coat or the gloves) inside the BSC i die a little bit inside. especially when they do it while talking about aseptic practice.
See I'm wondering if it's yet possible to split two cells from two different species and combine them with success? There is a cell that eats away at plastic. Could it be possible to combine it with another cell or organism?