Equivalency of Nuclear Transfer‐Derived Embryonic Stem Cells to Those Derived from Fertilized Mouse Blastocysts

Abstract

ABSTRACT

Establishment of customized mouse stem cell lines by sequential nuclear transfer

“…Karyotype analysis was thus performed for the different ES cell lines generated (Table 5) and percentages of normal karyotype in our ES cell lines appear to be similar to those reported previously [3].…”

“…NT-ESCs can be established successfully not only from cloned embryos with full developmental competency, but also from those with no potential for full-term development [3]. In this study, we generated 40 ESC lines from various embryos, including those derived from R1 ES cell donors at different passages, somatic donor cells (cumulus cells) and ES cells generated from the resulting somatic-clone embryos.…”

“…DNA was counterstained with Propidium Iodide (10 µg/ml) or/and 0.1 µg/ml Hoechst 3332 at room temperature for 5~10 min; Observation was performed under live cell station (Leica Co.) or confocal microscope (ZEISS, LSM 510 META). In some cell lines, the karyotype and EB formation was examined as described [3,26,27].…”

“…More recently, customized pluripotent embryonic stem cells (ESC) have been derived from nuclear transfer embryos by therapeutic cloning in several labs [2,3]. The ES cell lines generated were shown to be able to differentiate into all three embryonic germ layers both in vitro and in vivo, as well as form germlines leading to mature spermatozoa and oocytes [4][5][6][7][8][9][10].…”

“…A number of reports have shown the potential usefulness of therapeutic cloning using ES cells derived from cloned animals (NT-ESCs) in regenerative medicine to rescue immune deficient or degenerative phenotypes [12][13][14]. Other evidence suggested that NT-derived ES cells in mouse have similar developmental potential as fertilization-derived ES cells, and appear to be indistinguishable from their fertilization-derived counterparts on a molecular level [2,3].…”