Rederivation of transgenic mice from iPS cells derived from frozen tissue.
In mice, induced pluripotent stem (iPS) cells with embryonic stem (ES)-like characteristics have been derived by ectopic expression of four transcription factors in somatic cells: Sox2, Oct3/4, Klf4 and/or c-Myc. To date, iPS cells have only be made from freshly harvested tissues and cells. However, if iPS cells could be derived from frozen tissues and cells, then cryopreservation of tissues such as mouse tails could conceivably become a reliable alternative to the more traditional formats, like germplasm and ES cells, for the archiving of genetically altered mouse lines. To test this hypothesis, we sought to demonstrate that a live transgenic mouse line could be recovered from transgenic iPS cells derived from cryopreserved mouse tissues. Tails and tail-derived fibroblasts from a DsRED transgenic mouse were cryopreserved in the presence of 5% dimethylsulfoxide (DMSO) in liquid nitrogen for 1 week and 1 month, respectively. Afterward, tissues and cells were thawed and underwent nuclear reprogramming by molecular transfection to derive iPS cells which generated germline confirmed transgenic mice. Our results demonstrate for the first time that iPS cells can be efficiently derived from frozen-stored-thawed tail tissue and fibroblasts and used to re-establish a transgenic mouse line. Therefore, this study provides conclusive evidence that, as a practical matter, frozen tails and fibroblasts can be used as an effective and reliable alternative to frozen germplasm and ES cells for the storage, maintenance, and distribution of genetically-altered mutant mice.
Lee
Angus Yiu-Fai
AY
Mouse Biology Program, Center for Comparative Medicine, School of Veterinary Medicine, University of California, 2795 2nd Street, Suite 400, Davis, CA 95618, USA. ayflee@ucdavis.edu
Lloyd
K C Kent
KC
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051379:Mice; D008822:Mice, Transgenic; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D013623:Tail / Q000166:cytology*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014162:Transfection
C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D050814:Octamer Transcription Factor-3; D014157:Transcription Factors
Nature. 2009 Jul 2;460(7251):53-9
19483674
Cell. 2006 Aug 25;126(4):663-76
16904174
Proc Natl Acad Sci U S A. 1978 Nov;75(11):5565-9
281705
Curr Opin Genet Dev. 2001 Oct;11(5):587-94
11532403
Nature. 2009 Sep 3;461(7260):91-4
19672243
Science. 2008 Aug 1;321(5889):699-702
18276851
Nature. 1959 May 16;183(4672):1394-5
13657132
Proc Natl Acad Sci U S A. 1981 Dec;78(12):7634-8
6950406
Nature. 2009 Sep 3;461(7260):86-90
19672241
Circ Res. 2002 Aug 9;91(3):189-201
12169644
Nature. 2007 Jul 19;448(7151):313-7
17554338
Biol Reprod. 1993 Sep;49(3):489-95
8399840
Cell Stem Cell. 2008 Jan 10;2(1):10-2
18371415
Nat Biotechnol. 2008 Jan;26(1):101-6
18059259
Nature. 2007 Jul 19;448(7151):318-24
17554336
Nature. 1981 Jul 9;292(5819):154-6
7242681
Proc Natl Acad Sci U S A. 2008 Jul 8;105(27):9290-5
18579772
Nat Methods. 2009 May;6(5):363-9
19337237
Exp Cell Res. 1984 May;152(1):212-9
6714319
Stem Cells. 2009 Dec;27(12):2969-78
19816951
J Vet Med Sci. 2001 Oct;63(10):1151-4
11714036
Genesis. 2004 Dec;40(4):241-6
15593332
Cell Stem Cell. 2009 Aug 7;5(2):135-8
19631602