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First published online May 5, 2004
doi: 10.1242/10.1242/dev.01118

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The first cell cycle after transfer of somatic cell nuclei in a non-human primate

Soon-Chye Ng^*,, Naiqing Chen^*, Wan-Yue Yip^*, Swee-Lian Liow^*, Guo-Qing Tong, Barbara Martelli, Lay Geok Tan and Paolo Martelli

Laboratory of Nuclear Reprogramming and Cell Differentiation, Department of Obstetrics and Gynaecology, National University of Singapore, 5 Lower Kent Ridge Road, Singapore 119074, Singapore

View larger version (60K): [in a new window]   Fig. 1. PCC spindle formation in a SCNT-reconstructed LoTM zygote, 2 hours after injection of the donor LoTM cumulus cell. Red, DNA; green, -tubulin in confocal images. (A) Chromosome condensation and microtubule assembly into a normal PCC spindle. (i) Chromosomes in the middle of the PCC spindle. (ii) Another reconstructed zygote with chromosomes in the middle of the PCC spindle. (iii) Polarized microscopy view of a normal PCC spindle. (B) Chromosome condensation but improper capture onto normally assembled microtubules. (C) Chromosome condensation and microtubule assembly into an abnormal PCC spindle. (D) Chromosome condensation without microtubule assembly. (E) Non-condensed chromosomes and microtubule assembly into an abnormal PCC spindle. (F) No chromosome change and no microtubule assembly (arrow shows first polar body). (G) Normal separation in anaphase of a SCNT-reconstructed LoTM zygote, 2 hours after activation. (H) Abnormal separation of chromosomes in multiple directions on a diffused microtubule assembly, 2 hours after activation.

View larger version (114K): [in a new window]   Fig. 2. Results obtained following SCNT into enucleated LoTM oocytes. (A) Nuclear formation in a SCNT-reconstructed LoTM zygote, 14-16 hrs after activation. (i) Two nuclei, (ii) more than two nuclei, (iii) one nucleus and (iv) no nucleus. (B) Reconstructed embryos after the first cleavage, showing: (i) normal cleavage; (ii) abnormal cleavage resulting in fragmentation (iia) and/or unequal blastomeres (iib); and (iii) highly fragmented nuclei. (C) Pregnancy sacs on ultrasound for 99B7 on the 15th day (29 days after OR), OFC7 on the 15th day (29 days after OR), and 168D the 60th day (75 days after OR) after completion of the luteal phase.

View larger version (94K): [in a new window]   Fig. 3. Schematic of cell cycle changes following SCNT. (A) First cell cycle in reconstructed embryos following injection of somatic cells, with various possible pathways, with the number of embryos observed in parenthesis. (B) Chromosomes changes after nuclear reformation during the first cell cycle.

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