可信的大鼠胚胎干细胞的获取（英文)

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Capture of Authentic Embryonic Stem Cells from Rat Blastocysts
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2 k8 j7 `9 I9 JThis article is not included in your organization's subscription. However, you may be able to access this article under your organization's agreement with Elsevier.4 Z  x( ]% N4 _
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Mia Buehr1, 2, Stephen Meek1, 2, Kate Blair3, 4, Jian Yang3, 5, Janice Ure1, Jose Silva3, 4, Renee McLay1, John Hall3, 4, Qi-Long Ying1, 6 and Austin Smith3, 4, ,
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1Institute for Stem Cell Research, University of Edinburgh, King's Buildings, Edinburgh EH9 3JQ, UK$ K/ D7 [& ?! L* t' k
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2The Roslin Institute, University of Edinburgh, Roslin, Midlothian, EH25 9PS, UK
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4 V' C; h2 I) V2 a8 p3Wellcome Trust Centre for Stem Cell Research, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK
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4Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QW, UK
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5The Key Laboratory of Molecular Medicine, Ministry of Education, Shanghai Medical College, Fudan University, Shanghai, 200032, P.R. China
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Received 2 September 2008; + l3 W" V3 d6 B' G6 \. Y& d
revised 24 November 2008;
/ }  b! P0 h9 P, Raccepted 8 December 2008.
* F5 @4 p' l: t# V$ jPublished: December 24, 2008. . |. x( J% l% T9 A
Available online 24 December 2008.
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Refers to:  Germline Competent Embryonic Stem Cells Derived from Rat Blastocysts
2 e, |. f* H/ nCell, Volume 135, Issue 7, 26 December 2008, Pages 1299-1310,+ p8 p/ c; L: {+ N& N1 e/ [5 E
Ping Li, Chang Tong, Ruty Mehrian-Shai, Li Jia, Nancy Wu, Youzhen Yan, Robert E. Maxson, Eric N. Schulze, Houyan Song, Chih-Lin Hsieh, Martin F. Pera, Qi-Long Ying
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: `7 Z; f* t5 i3 q8 j3 O" rReferred to by:  Germline Competent Embryonic Stem Cells Derived from Rat Blastocysts; i8 U8 p9 j6 {3 L4 q" l' O9 a) @
Cell, Volume 135, Issue 7, 26 December 2008, Pages 1299-1310,
. x! |5 }. O7 ]) k) Z% h) ^Ping Li, Chang Tong, Ruty Mehrian-Shai, Li Jia, Nancy Wu, Youzhen Yan, Robert E. Maxson, Eric N. Schulze, Houyan Song, Chih-Lin Hsieh, Martin F. Pera, Qi-Long Ying4 z; D6 r9 {" @
Supplementary Content
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Embryonic stem (ES) cells have been available from inbred mice since 1981 but have not been validated for other rodents. Failure to establish ES cells from a range of mammals challenges the identity of cultivated stem cells and our understanding of the pluripotent state. Here we investigated derivation of ES cells from the rat. We applied molecularly defined conditions designed to shield the ground state of authentic pluripotency from inductive differentiation stimuli. Undifferentiated cell lines developed that exhibited diagnostic features of ES cells including colonization of multiple tissues in viable chimeras. Definitive ES cell status was established by transmission of the cell line genome to offspring. Derivation of germline-competent ES cells from the rat paves the way to targeted genetic manipulation in this valuable biomedical model species. Rat ES cells will also provide a refined test-bed for functional evaluation of pluripotent stem cell-derived tissue repair and regeneration.- H$ @: f( r. Z. I0 C* \
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Author Keywords: STEMCELL: ~# p# {+ \2 E& O  z0 ?9 E
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Article Outline
5 y9 z& m  T2 |' Z* X0 uIntroduction / }, c% L: y5 d: I7 A
Results * \& x+ e7 W* t$ [6 s8 [* Q
Sustained Expression of Pluripotent Markers in Rat Inner Cell Masses Explanted in 3i   [* o$ Q( M+ w( x# V1 c& l
Derivation of Continuous Cell Lines
4 X/ h8 f& ]* ]1 _6 ~Characterization of 3i Rat Cell Lines 9 g7 g  M. S. U" L' Q
Rat 3i Cells Contribute to Chimeras
4 k! ?3 F" M0 t( l2 }: `Derivation of Germline-Competent Rat Cells Using Two Inhibitors
/ {9 y: d$ K! Q0 |$ e" H0 }2i Rat Cells Exhibit In Vitro Properties and Behavior of Embryonic Stem Cells
7 k% l+ G3 }- f0 g+ X0 XDiscussion
- |% u7 R" s8 Z5 m+ }Experimental Procedures
+ y# k, _- h$ I3 ACulture Procedure * Q6 G- o* C6 f- R( I) D+ F8 ?0 j
Chimera Generation
6 N; L5 r' e! g5 N# i0 g  SImmunosurgery
  \, J9 O3 _/ ?' d" I& K2 O0 \0 ITeratoma Generation
/ D! W5 N  M; WElectroporation + P# K! a: F  H! ~
Cell-Cycle Profiling
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