原核移殖，冷凍與電融合後孤雌生殖分裂

將原核(germinal vesicle )之細胞核取出至移入去核之卵細胞中 再予以電融合後施行孤雌生殖分裂。
原核之細胞核亦可玻璃化冷凍儲存於去細胞質之中空透明帶(Zona pellucidae)中。
經冷凍解凍之原核可達到類似新鮮原核之生殖分裂。

http://www.jstage.jst.go.jp/article/jrd/57/3/335/_pdf

J Reprod Dev. 2011 Jun;57(3):335-41. Epub 2011 Jan 28.

Vitrification of fully grown and growing porcine oocytes using germinal vesicle transfer.

Nakagawa S, Maedomari N, Kikuchi K, Nagai T, Miyano T, Fulka J Jr, Manabe N.

Source

Animal Resource Science Center, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Kasama, Japan.

Abstract

The survival rate of vitrified germinal vesicle (GV) stage porcine oocytes is very low, and it is not known if the vitrificationdamages the nucleus, cytoplasm or both. We have evaluated the eventual GV or cytoplasmic damage in fully grown (FG) and growing vitrified oocytes. Fifty-five percent of nonvitrified FG cumulus-denuded oocytes reached the metaphase II (MII) stage in culture. When growing oocytes from preantral (PA) and early antral (EA) follicles were matured in vitro, almost all oocytes were arrested at the GV stage (GV stage: PA 88.9 and EA 79.5%, respectively). When fresh GVs from FG, PA and EA oocytes were transferred into fresh enucleated FG oocytes and matured in vitro, some of them reached the MII stage (MII stage: FG/FG 57.5%, PA/FG 9.3% and EA/FG 35.3%, respectively). The maturation rate of vitrified FG oocytes was only 6.1% but increased dramatically when vitrified GVs from FG, PA and EA oocytes were transferred into fresh enucleated FG oocytes (MII stage: VitFG/FG 43.9%, VitPA/FG 7.1% and VitEA/FG 26.3%, respectively). These results were not significantly different from those for the nonvitrified groups (MII stage: FG/FG 57.5%, PA/FG 9.3% and EA/FG 35.3%, respectively). We activated the reconstructed oocytes that received fresh or vitrified GVs (FG/FG, EA/FG, VitFG/FG and VitEA/FG) and examined their embryonic development. Cleaved embryos (nonvitrified groups 13.0-61.8%, vitrified groups 33.3-40.0%) and blastocysts (nonvitrified groups 0.0-18.2%, vitrified groups 0.0-2.9%) were obtained after activation. These results demonstrate that vitrified porcine GVs maintain maturational and developmental competence and that vitrificationpredominantly damages the cytoplasm.