哺乳動物細胞をトリプシン処理し、継代培養

Biology

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Summary

細胞がコンフルエントに達するように、彼らは、継代培養や継代する必要があります。このビデオでは、接着および浮遊細胞の両方を継代培養するための手順をデモンストレーションを行います。

Cite this Article

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Ricardo, R., Phelan, K. Trypsinizing and Subculturing Mammalian Cells. J. Vis. Exp. (16), e755, doi:10.3791/755 (2008).

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Abstract

細胞がコンフルエントに達するように、彼らは、継代培養や継代する必要があります。最終的には減少分裂指数と細胞死の中のサブカルチャーコンフルエントの細胞の結果に障害が発生。継代培養の最初のステップは、トリプシン処理や機械的手段によって初代培養容器の表面から細胞を剥離することです。結果として細胞懸濁液は、細分化、または新鮮な培養液に、再接種される。二次培養物は定期的に成長し、供給のためにチェックされ、続いて三次の文化を生成するために継代培養することができます。細胞の継代の間の時間は細胞株によって変化し、成長率に依存します。

Protocol

この実験的なアプローチのための完全なテキストのプロトコルでは、利用可能です細胞生物学におけるカレントプロトコール

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Comments

13 Comments

  1. the video isnot playing

    Reply
    Posted by: Anonymous
    July 6, 2008 - 10:23 PM
  2. how to prepare cell growth curve of  cancer cell line

    Reply
    Posted by: Anonymous
    September 11, 2008 - 3:10 AM
  3. can i know the passaging protocol for MDA MB ²31 Breast cancer cells.

    Reply
    Posted by: Anonymous
    September 18, 2008 - 12:10 PM
  4. sir can u tell me that how to transfer cell ( frozen into liq. nitrogen) into media and that will be the mother culture so in that also we have to do passaging , means we have to add HBSS and Trypsin and EDTA. and how to count cells. thank you waiting for your reply.... sam A Masih

    Reply
    Posted by: Anonymous
    September 24, 2008 - 12:24 PM
  5. Hi, For subculturing suspension cells, what is the normal routine? is it better if changed everyday or only after the media contents turns trubid and yellowish colour? Regards Kamal

    Reply
    Posted by: Anonymous
    October 22, 2008 - 7:45 PM
  6. Hello kamal bro..
    paras sir le bhannu bhayeko ta tyahi ho kya re...
    hemanta

    Reply
    Posted by: Anonymous
    July 28, 2009 - 7:18 AM
  7. Thank you

    Reply
    Posted by: Anonymous
    March 24, 2009 - 4:09 PM
  8. Could you elaborate on how to culture cancer cells? The precautions to be taken and when to subculture them. Thank you

    Reply
    Posted by: Preethi S.
    April 2, 2009 - 12:37 PM
  9. I can see the video

    Reply
    Posted by: Anonymous
    June 22, 2009 - 12:25 PM
  10. We were having a technical issue when you posted your comment. Please let us know at support@jove.com if you are unable to view this video.

    Reply
    Posted by: Anonymous
    June 23, 2009 - 7:52 PM
  11. every time I trypsinize my BBe cells - they clump together (even I don't shake the flask at all..)
    how can I prevent it ? because the clumps are super strong and I can not brake them down again.. I do something wrong?
    I wash ²x with PBS (37C) and 5-15min trypsinize the cells and add RPMI 1640 media carefully to it and mix gently..

    can you help me?

    thank you
    AlexK

    Reply
    Posted by: Alexandra K.
    April 12, 2010 - 5:19 PM
  12. thank u

    Reply
    Posted by: Anonymous
    October 21, 2010 - 6:32 AM
  13. How long dŒs it take for an MCF-7 cell lines grow confluent? What's the best media for culturing this cell?
    Thanks!

    Reply
    Posted by: Anonymous
    September 22, 2011 - 11:44 AM

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