Metaphase II karyoplast transfer from human in-vitro matured oocytes to enuclueated mature oocytes Atsushi Tanaka, Motoi Nagayoshi, Shoichiro Awata,

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Presentation transcript:

Metaphase II karyoplast transfer from human in-vitro matured oocytes to enuclueated mature oocytes Atsushi Tanaka, Motoi Nagayoshi, Shoichiro Awata, Norio Himeno, Izumi Tanaka, Seiji Watanabe, Hiroshi Kusunoki Reproductive BioMedicine Online Volume 19, Issue 4, Pages 514-520 (October 2009) DOI: 10.1016/j.rbmo.2009.06.004 Copyright © 2009 Reproductive Healthcare Ltd, Duck End Farm, Dry Drayton, Cambridge CB23 8DB, UK Terms and Conditions

Figure 1 Human oocytes at the germinal vesicle (arrowhead, A), the metaphase I (arrow, B) and the metaphase II stages (arrows, C and D), (C) recipient (in-vitro matured), (D) donor (in-vivo matured and freshly ovulated) oocyte. All photographs were taken after removing the cumulus cells. Original magnification, 400×. Reproductive BioMedicine Online 2009 19, 514-520DOI: (10.1016/j.rbmo.2009.06.004) Copyright © 2009 Reproductive Healthcare Ltd, Duck End Farm, Dry Drayton, Cambridge CB23 8DB, UK Terms and Conditions

Figure 2 Removing a karyoplast from an in-vitro matured metaphase II (MII) human oocyte. Oocyte held with the MII chromosomes at the 1–2 o’clock position (A). Zona pellucida just above the MII chromosomes breached with a glass micro-needle (B). Aspiration of MII chromosomes surrounded by a minimum amount of cytoplasm (C). Karyoplast expelled into the surrounding medium (D). Arrows indicate MII chromosomes (C and D). Original magnification, 400×. Reproductive BioMedicine Online 2009 19, 514-520DOI: (10.1016/j.rbmo.2009.06.004) Copyright © 2009 Reproductive Healthcare Ltd, Duck End Farm, Dry Drayton, Cambridge CB23 8DB, UK Terms and Conditions

Figure 3 Inserting a karyoplast into the perivitelline space of an enucleated metaphase II human oocyte matured in vivo. Isolated karyoplast (A) was aspirated again into the pipette (B). The pipette was inserted (C) and the karyoplast transferred into the perivitelline space (D). Arrows indicate metaphase II chromosome. Original magnification, 400×. Reproductive BioMedicine Online 2009 19, 514-520DOI: (10.1016/j.rbmo.2009.06.004) Copyright © 2009 Reproductive Healthcare Ltd, Duck End Farm, Dry Drayton, Cambridge CB23 8DB, UK Terms and Conditions

Figure 4 Electrofusion between a karyoplast and an ooplasm in human. Arrows indicate metaphase II chromosomes. Grafted oocyte (A) is placed between two microelectrodes (B). Fusing (C) and fused (D) karyoplast–cytoplast complex are shown after an electrical stimulation. Original magnification, 400×. Reproductive BioMedicine Online 2009 19, 514-520DOI: (10.1016/j.rbmo.2009.06.004) Copyright © 2009 Reproductive Healthcare Ltd, Duck End Farm, Dry Drayton, Cambridge CB23 8DB, UK Terms and Conditions

Figure 5 Chromosome plate of in-vitro matured oocytes (left). The karyotype was normal (23, X), but pre-divisions of sister chromatids were found in three chromosomes, one in each of B, C and E groups (arrows). Reproductive BioMedicine Online 2009 19, 514-520DOI: (10.1016/j.rbmo.2009.06.004) Copyright © 2009 Reproductive Healthcare Ltd, Duck End Farm, Dry Drayton, Cambridge CB23 8DB, UK Terms and Conditions

Figure 6 Scattered oocyte stained with aceto-orcein. The first polar body was out of focus. Original magnification, 400×. Reproductive BioMedicine Online 2009 19, 514-520DOI: (10.1016/j.rbmo.2009.06.004) Copyright © 2009 Reproductive Healthcare Ltd, Duck End Farm, Dry Drayton, Cambridge CB23 8DB, UK Terms and Conditions

Figure 7 Karyotype of a blastomere derived from a reconstituted human oocyte. Reproductive BioMedicine Online 2009 19, 514-520DOI: (10.1016/j.rbmo.2009.06.004) Copyright © 2009 Reproductive Healthcare Ltd, Duck End Farm, Dry Drayton, Cambridge CB23 8DB, UK Terms and Conditions