Characterization of pluripotent stem cells

被引：122

作者：

Marti, Merce ^{[1
]}

Mulero, Lola ^{[1
]}

Pardo, Cristina ^{[1
]}

Morera, Cristina ^{[1
]}

Carrio, Meritxell ^{[1
]}

Laricchia-Robbio, Leopoldo ^{[1
]}

Rodriguez Esteban, Concepcion ^{[1
,2
]}

Izpisua Belmonte, Juan Carlos ^{[1
,2
]}

机构：

[1] Ctr Regenerat Med Barcelona, Barcelona, Spain

[2] Salk Inst Biol Studies, Gene Express Lab, La Jolla, CA 92037 USA

来源：

NATURE PROTOCOLS | 2013年 / 8卷 / 02期

关键词：

EXPRESSION; DIFFERENTIATION; GENERATION; LINES;

D O I：

10.1038/nprot.2012.154

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Characterization of pluripotent stem cells is required for the registration of stem cell lines and allows for an impartial and objective comparison of the results obtained when generating multiple lines. It is therefore crucial to establish specific, fast and reliable protocols to detect the hallmarks of pluripotency. Such protocols should include immunocytochemistry (takes 2 d), identification of the three germ layers in in vitro-derived embryoid bodies by immunocytochemistry (immunodetection takes 3 d) and detection of differentiation markers in in vivo-generated teratomas by immunohistochemistry (differentiation marker detection takes 4 d). Standardization of the immunodetection protocols used ensures minimun variations owing to the source, the animal species, the endogenous fluorescence or the inability to collect large amounts of cells, thereby yielding results as fast as possible without loss of quality. This protocol provides a description of all the immunodetection procedures necessary to characterize mouse and human stem cell lines in different circumstances.

引用

页码：223 / 253

页数：31

共 31 条

[1] Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes [J].

Aasen, Trond ;

Raya, Angel ;

Barrero, Maria J. ;

Garreta, Elena ;

Consiglio, Antonella ;

Gonzalez, Federico ;

Vassena, Rita ;

Bilic, Josipa ;

Pekarik, Vladimir ;

Tiscornia, Gustavo ;

Edel, Michael ;

Boue, Stephanie ;

Izpisua Belmonte, Juan Carlos .

NATURE BIOTECHNOLOGY, 2008, 26 (11) :1276-1284

[2] Characterization of human embryonic stem cell lines by the International Stem Cell Initiative [J].

Adewumi, Oluseun ;

Aflatoonian, Behrouz ;

Ahrlund-Richter, Lars ;

Amit, Michal ;

Andrews, Peter W. ;

Beighton, Gemma ;

Bello, Paul A. ;

Benvenisty, Nissim ;

Berry, Lorraine S. ;

Bevan, Simon ;

Blum, Barak ;

Brooking, Justin ;

Chen, Kevin G. ;

Choo, Andre B. H. ;

Churchill, Gary A. ;

Corbel, Marie ;

Damjanov, Ivan ;

Draper, Jon S. ;

Dvorak, Petr ;

Emanuelsson, Katarina ;

Fleck, Roland A. ;

Ford, Angela ;

Gertow, Karin ;

Gertsenstein, Marina ;

Gokhale, Paul J. ;

Hamilton, Rebecca S. ;

Hampl, Ales ;

Healy, Lyn E. ;

Hovatta, Outi ;

Hyllner, Johan ;

Imreh, Marta P. ;

Itskovitz-Eldor, Joseph ;

Jackson, Jamie ;

Johnson, Jacqueline L. ;

Jones, Mark ;

Kee, Kehkooi ;

King, Benjamin L. ;

Knowles, Barbara B. ;

Lako, Majlinda ;

Lebrin, Franck ;

Mallon, Barbara S. ;

Manning, Daisy ;

Mayshar, Yoav ;

Mckay, Ronald D. G. ;

Michalska, Anna E. ;

Mikkola, Milla ;

Mileikovsky, Masha ;

Minger, Stephen L. ;

Moore, Harry D. ;

Mummery, Christine L. .

NATURE BIOTECHNOLOGY, 2007, 25 (07) :803-816

[3] Derivation of human embryonic stem cells at the Center of Regenerative Medicine in Barcelona [J].

Aran, Begona ;

Rodriguez-Piza, Ignasi ;

Raya, Angel ;

Consiglio, Antonella ;

Munoz, Yolanda ;

Barri, Pere N. ;

Carlos Izpisua, Juan ;

Veiga, Anna .

IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, 2010, 46 (3-4) :356-366

[4]

Campos P.B., 2009, J. Vis. Exp, V31, pe1512

[5]

Carpenter M.K., 2004, HANDBOOK OF STEM CEL, V1, P407

[6] Oct-4 mRNA and protein expression during human preimplantation development [J].

Cauffman, G ;

Van de Velde, H ;

Liebaers, I ;

Van Steirteghem, A .

MOLECULAR HUMAN REPRODUCTION, 2005, 11 (03) :173-181

[7] POU5F1 isoforms show different expression patterns in human embryonic stem cells and preimplantation embryos [J].

Cauffman, Greet ;

Liebaers, Inge ;

Van Steirteghem, Andre ;

Van de Velde, Hilde .

STEM CELLS, 2006, 24 (12) :2685-2691

[8]

Cerdan C., 2007, Curr. Protoc. Stem Cell Biol, V3, p1D.2.1, DOI DOI 10.2.1-10.2.16

[9] The International Stem Cell Banking Initiative (ISCBI): raising standards to bank on [J].

Crook, Jeremy Micah ;

Hei, Derek ;

Stacey, Glyn .

IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, 2010, 46 (3-4) :169-172

[10] Alternative Splicing Produces Nanog Protein Variants with Different Capacities for Self-renewal and Pluripotency in Embryonic Stem Cells [J].

Das, Satyabrata ;

Jena, Snehalata ;

Levasseur, Dana N. .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2011, 286 (49) :42690-42703

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