Human embryonic stem cell lines derived from single blastomeres

Abstract

The derivation of human embryonic stem (hES) cells currently requires the destruction of ex utero embryos. A previous study in mice indicates that it might be possible to generate embryonic stem (ES) cells using a single-cell biopsy similar to that used in preimplantation genetic diagnosis (PGD), which does not interfere with the embryo's developmental potential. By growing the single blastomere overnight, the resulting cells could be used for both genetic testing and stem cell derivation without affecting the clinical outcome of the procedure. Here we report a series of ten separate experiments demonstrating that hES cells can be derived from single blastomeres. In this proof-of-principle study, multiple biopsies were taken from each embryo using micromanipulation techniques and none of the biopsied embryos were allowed to develop in culture. Nineteen ES-cell-like outgrowths and two stable hES cell lines were obtained. The latter hES cell lines maintained undifferentiated proliferation for more than eight months, and showed normal karyotype and expression of markers of pluripotency, including Oct-4, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, nanog and alkaline phosphatase. These cells retained the potential to form derivatives of all three embryonic germ layers both in vitro and in teratomas. The ability to create new stem cell lines and therapies without destroying embryos would address the ethical concerns of many, and allow the generation of matched tissue for children and siblings born from transferred PGD embryos.

Derivation of human embryonic stem cell lines from single blastomeres of low-quality embryos by direct plating

“…The following year, these authors derived hESC lines from single blastomeres of thawed 8-cell stage embryos by co-culture with an established fluorescently-labeled hESC line, with a success rate of 2 % [5]. In addition, the derivation rate was increased to 20 % by co-culturing the blastomeres with the parent embryo and the addition of laminin to the medium [6].…”

“…However, clinical applicability can only be established after intensive laboratory research delineates simple and safe derivation protocols for hESC lines. Previous studies [5][6][7][8] have described the derivation of hESC lines from single blastomeres of high-quality (HQ) embryos with sequential culture, which is somewhat complicated. Therefore, a more convenient derivation method is desired.…”

Stem-cell research: Never say die

“…In August 2006, Lanza and his co-authors published a paper 6 in Nature showing that a single cell could be plucked from an 8-10-cell human embryo and grown into stem cells. Lanza wanted to show that it was possible to derive stem cells without destroying the embryo, to sidestep ethical concerns.…”

The impact of European embryonic stem cell patent decisions on research strategies

“…7 Moreover, in such a context of legal clarification on patentability with stem cells, some argued they never thought it will be possible to obtain patent on hESC in case of embryo destruction. 11 It can also be considered these legal decisions have a limited impact in the lights of future technological developments: 12 it is still possible to obtain patents where hESC are obtained without embryo destruction 13 and IPS may provide new patent possibilities. While patents as such can be seen as necessary for investors, notably as they limit secrecy, patents' unavailability permits higher freedom of activities (no fear of patents' rights infringement, no royalties to pay for exploitative license) and other means to stimulate innovation can be used.…”