A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture (original) (raw)

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09:18 min

August 11th, 2014

DOI :

10.3791/51765-v

August 11th, 2014


Amanda Andersson-Rolf1,2, Juergen Fink1,2, Roxana C. Mustata2, Bon-Kyoung Koo1,2

1Department of Genetics, University of Cambridge, 2Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge

0:05

Title

1:17

Crypt Isolation

3:39

Virus Production

5:15

Retroviral Transduction

6:34

Spinoculation and Plating

8:01

Results: Representative Organoid Images

8:49

Conclusion

This protocol explains primary Lgr5-positve organoid culture and the subsequent performance of retroviral transduction. This enables Cre-inducible overexpression or knockdown of the delivered transgene and allows functional studies to be carried out in the novel in vitro organotypicmodel system_._

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