Introduction to cell culture, splitting cells using trypsin and counting them using a hemocytometer

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  • Опубліковано 1 вер 2020
  • 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.
  • Наука та технологія

КОМЕНТАРІ • 47

  • @bariatricbabe7438
    @bariatricbabe7438 Рік тому +15

    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.

    • @user-ur7gk8st3x
      @user-ur7gk8st3x 4 місяці тому

      Hello can you plss tell me application of immunohistochemistry

    • @funny11744
      @funny11744 Місяць тому

      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 ?

    • @kosheeka
      @kosheeka День тому

      @@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.

  • @paolarosado8348
    @paolarosado8348 3 роки тому +12

    Great video and explanations! Thank you for uploading it. I really enjoyed the camera technique, because is like you are doing it yourself.

  • @virgirma1328_PhD_scholar
    @virgirma1328_PhD_scholar Рік тому +5

    Very thorough tutorial !Thanks teacher !

  • @Lunedai
    @Lunedai 3 роки тому +2

    I am so greatful for this helpful video! Thank you so much

  • @MonikaShailesh777
    @MonikaShailesh777 3 роки тому +2

    Thank u Olga 💓...keep posting, really helpful video 😊

  • @pichej22
    @pichej22 Рік тому +1

    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.

  • @alessiadauria1490
    @alessiadauria1490 Рік тому

    Great video, thanks! Very helpul for my first times in the lab

  • @biologylover1565
    @biologylover1565 2 роки тому +1

    Amazing video, Thank you for your effort. 👍👍👍

  • @Koonnuusana
    @Koonnuusana 3 роки тому +1

    Thank you so much for this good video. It make me clearly understand about cell culture. 😊❤🍀👩‍🔬

  • @alissaa.2287
    @alissaa.2287 Рік тому

    You are a hero to me, thank you.

  • @ady12377
    @ady12377 14 днів тому

    This helped me alot! Thank you maam ! ❤ may you live a long and meaningful life ❤❤❤😊

  • @tseringsamkhar1198
    @tseringsamkhar1198 3 роки тому +1

    Very insightful . Thank u

  • @zuhayousuf9197
    @zuhayousuf9197 2 роки тому

    Such a good video! Thank you :)

  • @hallahadi5548
    @hallahadi5548 2 роки тому

    braaaaaava you are the best one who was explanation cell culture to me >>> thanks a lot

    •  Рік тому

      Happy to hear that!

  • @aydafathollahpour602
    @aydafathollahpour602 Рік тому

    Thank you, it was amazing

  • @shutterismmn
    @shutterismmn 2 роки тому

    this video made my concept clear. thanks

    •  Рік тому

      You're welcome!

  • @jacqueonassismec3430
    @jacqueonassismec3430 3 місяці тому

    Thank you very much for this teaching..

  • @ko-fanta9829
    @ko-fanta9829 3 місяці тому

    well prepared and clear

  • @JYOtiRaNJanMANgaRaj
    @JYOtiRaNJanMANgaRaj 3 місяці тому +1

    Thank you so much 🙏🙏🙏❤❤❤❤❤❤❤❤❤❤

  • @mdselimahamed8617
    @mdselimahamed8617 2 місяці тому

    clear explanation indeed .

  • @anoopyes5004
    @anoopyes5004 11 місяців тому

    Tysm for the video❤

  • @niloofarkh4779
    @niloofarkh4779 Рік тому

    THANK U SO MUCH

  • @curiouscreature747
    @curiouscreature747 2 роки тому

    thanks olga soto

  • @sainaboujatta3108
    @sainaboujatta3108 2 роки тому

    helpful!

  • @livinghope8039
    @livinghope8039 2 роки тому

    great job, I liked and sub.. it

  • @fallseason5163
    @fallseason5163 2 роки тому

    Super super thank you for sharing it with us… do you have experience with BREAST CANCER STEM CELLS???

  • @funny11744
    @funny11744 2 роки тому +1

    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

  • @osmirod1
    @osmirod1 Рік тому +1

    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?

  • @mxasf
    @mxasf Рік тому +2

    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.

    •  Рік тому +1

      Great point!

  • @SweetMangoMuncher
    @SweetMangoMuncher Рік тому +1

    More trypsin may be necessary depending on the potency of the trypsin as well

    • @kosheeka
      @kosheeka Місяць тому +1

      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.

  • @ruqaiyatasneem8594
    @ruqaiyatasneem8594 11 днів тому

    What about renoving the neutralized trypsin and adding fresh media ?

  • @dhivyadharshiniboopalan2069

    What is the name of that aspirator used in discarding ?

  • @samratpaul25
    @samratpaul25 3 роки тому +4

    Are any animals killed during the extraction of Trypsin from their pancreas ? Or can it be done without killing the animal ?

    •  2 роки тому +11

      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.

  • @amitshivhare6690
    @amitshivhare6690 3 місяці тому

    A̺m̺a̺z̺i̺n̺g̺ v̺i̺d̺e̺o̺ ❤

  • @monaelrefay2956
    @monaelrefay2956 Рік тому

    Do you make other videos for MTT assay

  • @handydan6273
    @handydan6273 3 роки тому

    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?

  • @getoverit2800
    @getoverit2800 3 роки тому +2

    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.

  • @funny11744
    @funny11744 Рік тому

    At the minute 0:47 it is written ,, Immortalized cells could ....indefinetely" . Somebody could explain ?

    • @kosheeka
      @kosheeka 4 години тому

      It says "reproduce indefinitely." It means when these immortalized cells are grown under favorable condition they can reproduce indefinitely.