Trypsinizing and Subculturing Mammalian Cells

doi:10.3791/755

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Trypsinizing and Subculturing Mammalian Cells

Richard Ricardo, Katy Phelan
Molecular Pathology Laboratory, Inc.


			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.


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


			Cite this Article

Ricardo R, Phelan K (2008). Trypsinizing and Subculturing Mammalian Cells. JoVE. 16. http://www.jove.com/index/details.stp?id=755, doi: 10.3791/755


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07/06/2008 10:23:10 PM

shirri responded with a statement of type: Neutral

the video isnot playing

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09/11/2008 3:10:05 AM

amit responded with a statement of type: Neutral

how to prepare cell growth curve of cancer cell line

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09/18/2008 12:10:47 PM

nav responded with a statement of type: Question

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

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09/24/2008 12:24:28 PM

sam responded with a statement of type: Neutral

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

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10/22/2008 7:45:39 PM

kamal prajapati responded with a statement of type: Question

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

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07/28/2009 7:18:12 AM

hemanta responded with a statement of type: Neutral

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

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03/24/2009 4:09:31 PM

Ammar responded with a statement of type: Neutral

Thank you

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04/02/2009 12:37:38 PM

preethi_p2 responded with a statement of type: Neutral

Could you elaborate on how to culture cancer cells? The precautions to be taken and when to subculture them.

Thank you

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06/22/2009 12:25:21 PM

RF responded with a statement of type: Neutral

I can see the video

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06/23/2009 7:52:18 PM

nikitab responded with a statement of type: Neutral

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.

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04/12/2010 5:19:28 PM

alexk responded with a statement of type: Question

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

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Keywords

Basic Protocols, Issue 16, Current Protocols Wiley, Cell Culture, Cell Passaging, Trypsinizing Cells, Adherent Cells, Suspension Cells

Info

06/12/2008

Publish Date

20166

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doi: 10.3791/755

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Journal of Visualized Experiments (JoVE) is an online research journal employing visualization to increase reproducibility and transparency in biological sciences.

ISSN 1940-087X

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