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포유 동물 세포를 Trypsinizing 및 Subculturing

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Molecular Pathology Laboratory Network, Inc

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Cite this Article: 포유 동물 세포를 Trypsinizing 및 Subculturing

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

Abstract: 포유 동물 세포를 Trypsinizing 및 Subculturing

세포 confluency 도달로, 그들은 subcultured 또는 passaged해야합니다. 결국 감소 mitotic 색인 및 세포 죽음에의 subculture 합류 전지 결과에 오류가 발생했습니다. subculturing의 첫 번째 단계는 trypsinization 또는 기계적 수단으로 기본 문화 선박의 표면에서 세포를 분리하는 것입니다. 결과 세포 현탁액은 다음 세분화, 또는 신선한 문화로, reseeded입니다. 차 문화가 성장을위한 확인하고 정기적으로 공급하고, 이후 차 문화를 생산하는 subcultured 수 있습니다. 세포의 passaging 사이의 시간은 세포주에 따라 다릅니다 및 성장 속도에 따라 다릅니다.

Protocol: 포유 동물 세포를 Trypsinizing 및 Subculturing

이 실험 방법에 대한 전체 텍스트 프로토콜에서 사용할 수 있습니다 셀 생물학의 현재 프로토콜 .

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Disclosures: 포유 동물 세포를 Trypsinizing 및 Subculturing

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11 Comments

the video isnot playing

1

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

how to prepare cell growth curve of  cancer cell line

2

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

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

4

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Posted by: navSeptember 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

5

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Posted by: samSeptember 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

6

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Posted by: kamal prajapatiOctober 22, 2008, 7:45 PM

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

6.1

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

Thank you

7

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Posted by: AmmarMarch 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

8

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Posted by: Preethi S.April 2, 2009, 12:37 PM

I can see the video

9

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Posted by: RFJune 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.

9.1

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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

10

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Posted by: Alexandra K.April 12, 2010, 5:19 PM

thank u

11

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Posted by: mohamed ragabOctober 21, 2010, 6:32 AM

How long does it take for an MCF-7 cell lines grow confluent? What's the best media for culturing this cell?
Thanks!

12

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Posted by: TinaSeptember 22, 2011, 11:44 AM

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