Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment

被引:212
作者
Rodin, Sergey [1 ]
Antonsson, Liselotte [2 ,3 ]
Niaudet, Colin [4 ]
Simonson, Oscar E. [5 ,6 ]
Salmela, Elina [7 ,8 ]
Hansson, Emil M. [9 ]
Domogatskaya, Anna [1 ]
Xiao, Zhijie [1 ]
Damdimopoulou, Pauliina [2 ,3 ]
Sheikhi, Mona [2 ,3 ]
Inzunza, Jose [2 ,3 ]
Nilsson, Ann-Sofie [1 ]
Baker, Duncan [10 ]
Kuiper, Raoul [11 ]
Sun, Yi [12 ]
Blennow, Elisabeth [13 ,14 ]
Nordenskjold, Magnus [13 ,14 ]
Grinnemo, Karl-Henrik [5 ,6 ]
Kere, Juha [7 ,8 ,15 ,16 ]
Betsholtz, Christer [4 ]
Hovatta, Outi [2 ,3 ]
Tryggvason, Karl [1 ,17 ]
机构
[1] Karolinska Inst, Dept Med Biochem & Biophys, Div Matrix Biol, SE-17177 Stockholm, Sweden
[2] Karolinska Inst, Dept Clin Sci Intervent & Technol, Div Obstet & Gynecol, SE-14186 Huddinge, Sweden
[3] Karolinska Univ Hosp, SE-14186 Huddinge, Sweden
[4] Karolinska Inst, Dept Med Biochem & Biophys, SE-17177 Stockholm, Sweden
[5] Karolinska Inst, Adv Ctr Translat Regenerat Med ACTREM, Dept Mol Med & Surg, Div Cardiothorac Surg & Anaesthesiol, SE-17177 Stockholm, Sweden
[6] Karolinska Univ Hosp, SE-17177 Stockholm, Sweden
[7] Univ Helsinki, Res Programs Unit, FI-00014 Helsinki, Finland
[8] Folkhalsan Inst Genet, FI-00014 Helsinki, Finland
[9] Massachusetts Gen Hosp, Cardiovasc Res Ctr, Boston, MA 02114 USA
[10] Sheffield Childrens NHS Trust, Sheffield Diagnost Genet Serv, Sheffield S10 2TH, S Yorkshire, England
[11] Karolinska Hosp, Karolinska Inst, FENO, Dept Lab Med, SE-14186 Stockholm, Sweden
[12] BioLamina AB, S-17266 Sundbyberg, Sweden
[13] Karolinska Univ Hosp, Dept Mol Med & Surg, S-17176 Stockholm, Sweden
[14] Karolinska Univ Hosp, Karolinska Inst, Ctr Mol Med, S-17176 Stockholm, Sweden
[15] Karolinska Inst, Dept Biosci & Nutr, SE-14183 Stockholm, Sweden
[16] Karolinska Inst, Sci Life Lab, SE-14183 Stockholm, Sweden
[17] Duke NUS, Cardiovasc & Metab Disorders Program, Singapore 169857, Singapore
来源
NATURE COMMUNICATIONS | 2014年 / 5卷
基金
英国医学研究理事会; 瑞典研究理事会;
关键词
TERM SELF-RENEWAL; E-CADHERIN; LINES; GROWTH; DERIVATION; EXPRESSION; EXPANSION; APOPTOSIS; SURVIVAL; ADHESION;
D O I
10.1038/ncomms4195
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Lack of robust methods for establishment and expansion of pluripotent human embryonic stem (hES) cells still hampers development of cell therapy. Laminins (LN) are a family of highly cell-type specific basement membrane proteins important for cell adhesion, differentiation, migration and phenotype stability. Here we produce and isolate a human recombinant LN-521 isoform and develop a cell culture matrix containing LN-521 and E-cadherin, which both localize to stem cell niches in vivo. This matrix allows clonal derivation, clonal survival and long-term self-renewal of hES cells under completely chemically defined and xeno-free conditions without ROCK inhibitors. Neither LN-521 nor E-cadherin alone enable clonal survival of hES cells. The LN-521/E-cadherin matrix allows hES cell line derivation from blastocyst inner cell mass and single blastomere cells without a need to destroy the embryo. This method can facilitate the generation of hES cell lines for development of different cell types for regenerative medicine purposes.
引用
收藏
页数:13
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