Open AccessDerivation of Human Embryonic Stem Cells from Day‐8 Blastocysts Recovered after Three‐Step In Vitro Culture
Author(s) -
Stojkovic Miodrag,
Lako Majlinda,
Stojkovic Petra,
Stewart Rebecca,
Przyborski Stefan,
Armstrong Lyle,
Evans Jerry,
Herbert Mary,
Hyslop Louise,
Ahmad Sajjad,
Murdoch Alison,
Strachan Tom
Publication year - 2004
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.22-5-790
Subject(s) - homeobox protein nanog , biology , inner cell mass , embryonic stem cell , stem cell , microbiology and biotechnology , kosr , cell culture , andrology , in vitro , alkaline phosphatase , karyotype , blastocyst , rex1 , cellular differentiation , immunology , embryogenesis , adult stem cell , embryo , induced pluripotent stem cell , genetics , gene , chromosome , biochemistry , medicine , enzyme
Human embryonic stem cells (hESCs) have been derived from the inner cell mass (ICM) of day 5–7 blastocysts and hold great promise for research into human developmental biology and the development of cell therapies for the treatment of human diseases. We report here that our novel three‐step culture conditions successfully support the development of day‐8 human blastocysts, which possess significantly ( p <.01) more ICM cells than day‐6 blastocysts. Plating of ICMs isolated from day‐8 blastocysts resulted in the formation of a colony with hESC morphology from which a new hESC line (hES‐NCL1) was derived. Our stem cell line is characterized by the expression of specific cell surface and gene markers: GTCM‐2, TG343, TRA1‐60, SSEA‐4, alkaline phosphatase, OCT‐4 , NANOG , and REX‐1 . Cytogenetic analysis of the hESCs revealed that hES‐NCL1 line has a normal female (46, XX) karyotype. The pluripotency of the cell line was confirmed by the formation of teratomas after injection into severely combined immunodeficient mice and spontaneous differentiation under in vitro conditions.