JoVE   
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Biology

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Neuroscience

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Immunology and Infection

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Clinical and Translational Medicine

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Bioengineering

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Applied Physics

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Chemistry

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Behavior

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Environment

|   

JoVE Science Education

General Laboratory Techniques

You do not have subscription access to videos in this collection. Learn more about access.

Basic Methods in Cellular and Molecular Biology

You do not have subscription access to videos in this collection. Learn more about access.

Model Organisms I

You do not have subscription access to videos in this collection. Learn more about access.

Model Organisms II

You do not have subscription access to videos in this collection. Learn more about access.

 JoVE Biology

Trypsinizing and Subculturing Mammalian Cells

1, 1

1Molecular Pathology Laboratory Network, Inc

Article
    Downloads Comments Metrics

    You must be subscribed to JoVE to access this content.

    This article is a part of   JoVE Biology. If you think this article would be useful for your research, please recommend JoVE to your institution's librarian.

    Recommend JoVE to Your Librarian

    Current Access Through Your IP Address

    You do not have access to any JoVE content through your current IP address.

    IP: 54.235.5.178, User IP: 54.235.5.178, User IP Hex: 921372082

    Current Access Through Your Registered Email Address

    You aren't signed into JoVE. If your institution subscribes to JoVE, please or create an account with your institutional email address to access this content.

     

    Summary

    As cells reach confluency, they must be subcultured or passaged. This video will demonstrate a procedure for subculturing both adherent and suspension cells.

    Date Published: 6/12/2008, Issue 16; doi: 10.3791/755

    Cite this Article

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

    Abstract

    As cells reach confluency, they must be subcultured or passaged. Failure to subculture confluent cells results in reduced mitotic index and eventually in cell death. The first step in subculturing is to detach cells from the surface of the primary culture vessel by trypsinization or mechanical means. The resultant cell suspension is then subdivided, or reseeded, into fresh cultures. Secondary cultures are checked for growth and fed periodically, and may be subsequently subcultured to produce tertiary cultures. The time between passaging of cells varies with the cell line and depends on the growth rate.

    Protocol

    The complete text protocol for this experimental approach is available in Current Protocols in Cell Biology.

    Subscription Required. Please recommend JoVE to your librarian.

    Disclosures

    The authors have nothing to disclose.

    Comments

    13 Comments

    the video isnot playing
    Reply

    Posted by: AnonymousJuly 6, 2008, 10:23 PM

    how to prepare cell growth curve of  cancer cell line
    Reply

    Posted by: AnonymousSeptember 11, 2008, 3:10 AM

    can i know the passaging protocol for MDA MB ²31 Breast cancer cells.
    Reply

    Posted by: AnonymousSeptember 18, 2008, 12:10 PM

    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: AnonymousSeptember 24, 2008, 12:24 PM

    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: AnonymousOctober 22, 2008, 7:45 PM

    Hello kamal bro..
    paras sir le bhannu bhayeko ta tyahi ho kya re...
    hemanta
    Reply

    Posted by: AnonymousJuly 28, 2009, 7:18 AM

    Thank you
    Reply

    Posted by: AnonymousMarch 24, 2009, 4:09 PM

    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

    I can see the video
    Reply

    Posted by: AnonymousJune 22, 2009, 12:25 PM

    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: AnonymousJune 23, 2009, 7:52 PM

    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

    thank u
    Reply

    Posted by: AnonymousOctober 21, 2010, 6:32 AM

    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: AnonymousSeptember 22, 2011, 11:44 AM

    Post a Question / Comment / Request

    You must be signed in to post a comment. Please or create an account.

    Metrics

    Waiting
    simple hit counter