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Merck
CN

Derivation of human embryonic stem cells from single blastomeres.

Nature protocols (2007-08-19)
Irina Klimanskaya, Young Chung, Sandy Becker, Shi-Jiang Lu, Robert Lanza
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Laminin from human placenta, liquid, BioReagent, suitable for cell culture
Sigma-Aldrich
Leukemia Inhibitory Factor human, 10 µg, human recombinant LIF protein, expressed in E. coli, suitable for stem cell culture
Sigma-Aldrich
Fibronectin human plasma, lyophilized powder, BioReagent, suitable for cell culture