Derivation of human embryonic stem cells from single blastomeres.
This protocol details a method to derive human embryonic stem (hES) cells from single blastomeres. Blastomeres are removed from morula (eight-cell)-stage embryos and cultured until they form multicell aggregates. These blastomere-derived cell aggregates are plated into microdrops seeded with mitotically inactivated feeder cells, and then connected with neighboring microdrops seeded with green fluorescent protein-positive hES cells. The resulting blastomere-derived outgrowths are cultured in the same manner as blastocyst-derived hES cells. The whole process takes about 3-4 months.
Klimanskaya
Irina
I
Advanced Cell Technology, Worcester, Massachusetts 01605, USA.
Chung
Young
Y
Becker
Sandy
S
Lu
Shi-Jiang
SJ
Lanza
Robert
R
D000818:Animals; D001757:Blastomeres / Q000166:cytology* / Q000187:drug effects; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000166:cytology*; D005347:Fibroblasts / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000032:analysis; D006801:Humans; D051379:Mice; D016685:Mitomycin / Q000494:pharmacology
D049452:Green Fluorescent Proteins; D016685:Mitomycin