caa a22 4500 445291931 CHVBK 20180317142530.0 cr unu---uuuuu 170323e20100401xx s 000 0 eng 10.1007/s11626-010-9280-8 doi (NATIONALLICENCE)springer-10.1007/s11626-010-9280-8 Derivation of a new human embryonic stem cell line, Endeavour-2, and its characterization [Elektronische Daten] [Kuldip Sidhu, John Ryan, Justin Lees, Bernard Tuch] Here, we describe the derivation of a novel human embryonic stem cell (hESC) line, Endeavour-2 (E-2), propagated on human fetal fibroblasts (HFF) in a serum-replacement media. The inner cell mass (ICM) was manually dissected from the blastocyst without using immunodissection and, therefore, antibodies from animal sources. A total of 20 embryos were thawed and cultured, eight embryos were hatched, and five ICMs were obtained. They were transferred onto HFF used as feeder layer, and one colony representing the initial cell proliferation of a new hESC line, E-2, was obtained. The newly emerged hESC colony was passaged first by physical dissection and subsequently by enzymatic dissociation. E-2 has been in culture for over 6months and has been shown to possess typical features of a pluripotent hESC line including expression of stem cell surface markers (SSEA4, TRA-160, and integrin alpha-6), intracellular alkaline phosphatase, and pluripotency gene markers, OCT4 and NANOG. This hESC line shows lineage-specific differentiation into various representative cell types expressing markers characteristic of the three somatic germ layers under both in vitro and in vivo conditions. E-2 line shows a normal karyotype (46 XX) and has been successfully cryopreserved and thawed several times using slow-freezing procedures. E-2 adds to the repertoire of existing hESC lines for research and development purposes in the field of regenerative medicine. The Society for In Vitro Biology, 2010 Pluripotency nationallicence Teratomas nationallicence Blastocyst nationallicence Differentiation nationallicence Sidhu Kuldip Diabetes Transplant Unit, Prince of Wales Hospital & University of New South Wales, Sydney, NSW, Australia aut Ryan John IVF Australia, Sydney, NSW, Australia aut Lees Justin Diabetes Transplant Unit, Prince of Wales Hospital & University of New South Wales, Sydney, NSW, Australia aut Tuch Bernard Diabetes Transplant Unit, Prince of Wales Hospital & University of New South Wales, Sydney, NSW, Australia aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/3-4(2010-04-01), 269-275 1071-2690 46:3-4<269 2010 46 11626 https://doi.org/10.1007/s11626-010-9280-8 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11626-010-9280-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sidhu Kuldip Diabetes Transplant Unit, Prince of Wales Hospital & University of New South Wales, Sydney, NSW, Australia aut NATIONALLICENCE 700 1- Ryan John IVF Australia, Sydney, NSW, Australia aut NATIONALLICENCE 700 1- Lees Justin Diabetes Transplant Unit, Prince of Wales Hospital & University of New South Wales, Sydney, NSW, Australia aut NATIONALLICENCE 700 1- Tuch Bernard Diabetes Transplant Unit, Prince of Wales Hospital & University of New South Wales, Sydney, NSW, Australia aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/3-4(2010-04-01), 269-275 1071-2690 46:3-4<269 2010 46 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer