Improvement of canine somatic cell nuclear transfer procedure

Jang, Goo; Oh, Hyun Ju; Kim, M.K.; Fibrianto, Yuda Heru; Hossein, Mohammad Shamim; Kim, H.J.; Kim, J.J.; Hong, So Gun; Park, Jung E.; Kang, Sa-Ouk; Lee, B.C.

doi:10.1016/j.theriogenology.2007.08.022

Cited by 26 publications

(18 citation statements)

References 35 publications

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“…However, SCNT embryos did not progress to advanced stages (morula or blastocyst) and the percentage was very low even in the parthenogenetic group. These results correspond with the previous canine studies (Jang et al, 2008;Rodrigues Bde et al, 2004Songsasen et al, 2002), suggesting that both the in vitro culture system and the SCNT procedure would preclude a long-term preimplantation development in dogs. Concerning this issue, embryo transfer was done at the one-cell stage as previously reported (Jang et al, 2008).…”

Section: Discussionsupporting

confidence: 91%

“…Successful enucleation was confirmed by nuclear staining and a donor somatic cell was transferred to the perivitelline space of the recipient oocyte by previously reported procedure (Jang et al, , 2008. After microinjection, a couplet of donor cell-cytoplast complexes were induced to fuse with two pulses of direct current of 72 V cm 21 for 15 ls each using an Electro-Cell Fusion apparatus (NEPA GENE, Chiba, Japan).…”

Section: Collection Of In Vivo Matured Oocyte and Scntmentioning

confidence: 99%

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Generation of red fluorescent protein transgenic dogs

Hong

Kim

Jang

et al. 2009

Genesis

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Dogs (Canis familiaris) share many common genetic diseases with humans and development of disease models using a transgenic approach has long been awaited. However, due to the technical difficulty in obtaining fertilizable eggs and the unavailability of embryonic stem cells, no transgenic dog has been generated. Canine fetal fibroblasts were stably transfected with a red fluorescent protein (RFP) gene-expressing construct using retrovirus gene delivery method. Somatic cell nuclear transfer was then employed to replace the nucleus of an oocyte with the nucleus of the RFP-fibroblasts. Using this approach, we produced the first generation of transgenic dogs with four female and two male expressing RFP.

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Section: Discussionsupporting

confidence: 91%

Section: Collection Of In Vivo Matured Oocyte and Scntmentioning

confidence: 99%

Generation of red fluorescent protein transgenic dogs

Hong

Kim

Jang

et al. 2009

Genesis

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“…Une seule étude rapporte l'utilisation de cette technique pour la maturation nucléaire d'ovocytes canins immatures, le taux de reprise de la méiose ayant été de 5,9 % au bout de quatre jours (Lee et al 2005). C'est également par cette technique que les embryons issus du clonage de cellules somatiques de chien (donc au stade d'une cellule) ont été transplantés in utero chez la chienne dès leur reconstitution (Jang et al 2008 …”

Section: Transplantations Homospécifiquesunclassified

Microenvironnement tubaire: rôle dans la maturation des ovocytes canins in vivo et in vitro

Fontaine¹,

Reynaud²,

Thoumire³

et al. 2009

bavf

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Chez la plupart des mammifères, les ovocytes sont bloqués en métaphase II au moment de l'ovulation et cette inhibition de la méiose est ensuite levée par la fécondation. Chez les chiennes et les autres femelles de canidés, les ovocytes sont libérés au stade de prophase I, et il faut encore attendre 48 à 72 heures pour qu'ils atteignent le stade de métaphase II et deviennent fécondables. Cette particularité constitue aujourd'hui un frein au développement des biotechnologies de la reproduction chez les canidés. En effet, dans les essais de maturation in vitro d'ovocytes canins, seuls 10 à 30 % des ovocytes atteignent le stade de métaphase au bout de 72 heures de culture. Chez la chienne, la maturation nucléaire se produisant dans l'oviducte, des substituts de l'oviducte (milieux de culture comme le Synthetic Oviductal Fluid, explants d'oviductes, cultures sur tapis de cellules tubaires) ont été utilisés pour les cultures d'ovocytes in vitro, afin d'en améliorer le rendement, mais sans grand succès jusqu'à maintenant. Cet échec peut être dû au manque de données sur la composition du liquide tubaire de la chienne. L'étude de ce microenvironnement prend donc tout son intérêt, celui-ci étant probablement assez différent de celui des autres femelles, ne serait-ce que par l'existence du processus de lutéinisation préovulatoire dans cette espèce. À terme, la conception d'un milieu de maturation sur la base de la composition du liquide tubaire pourrait être une voie intéressante pour augmenter les taux de maturation in vitro. Mots

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“…Following development of cloned embryos directly depends on successful association of the host and donor cells, and viability of the resulting hybrid (10)(11)(12)(13)(14).…”

mentioning

confidence: 99%

Optimization of Parameters of the Oocyte Enucleation and Fusion With the Somatic Cell During Production of Mammalian Cloned Embryos

Сингина¹,

Лопухов²,

Zinovieva³

et al. 2013

S-h. biol.

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S u m m a r yParameters of the oocyte enucleation and fusion with somatic cell for producing of cattle cloned cetohybrids were studied. It was established that the electrofusion efficiency depends on strength, duration and set number of electrical pulse. The optimal parameters for Multiporator (Eppendorf, Germany) were determined: two consistent pulses DC electrical field each of 30 V field strength and 15 msec duration followed by the pulse of AC electrical field of 5 V and 10 sec duration. It was found that the first polar body serving as a marker for the chromosome position at «blind» enucleation must not deviate from the chromosome location by more than 10 degrees.Keywords: in vitro culture, oocyte, cattle nuclear transfer, electrofusion, enucleation.Somatic cloning is widely used in biomedical research and in production of transgenic animals (1). Despite a certain success of using this technique in recent years, it is yet inefficient in cattle and other domestic animals owing to high incidence of embryonic abnormalities and low viability of the born cloned offspring (2-4).To create a clone, genetic material of the host's egg cell is replaced by genetic material of a somatic cell (reprogramming). It is often associated with complete removal of the oocyte's chromosomes at metaphase II, after which the somatic donor cell is introduced in the perivitelline space of the (5).The procedure is visualized by staining of the chromosomes with fluorescent vital dye Hoechst 33342 allowing enucleation of the host oocyte under ultraviolet light (6). However, this approach adversely affects viability of cells and future embryos (7). The alternative solution is blind enucleation when metaphase chromosomes are recognized by location of the first polar body that serves as a reference point. This method is considered as more safe for viability of the egg cells and their capacity to future embryonic development (8), though some problems are not excluded too. After isolation of the first spindle pole body, metaphase plate of chromosomes stays for a while in a close proximity to it. Then, with aging of the egg cell, chromosomes disconnect from the spindle pole body that migrates to the perivitelline space. This circumstance complicates blind enucleation of oocytes or interdicts it (9), which necessitates identifying the factors important for efficiency of the method.After enucleation, the fusion of cytoplast and karyoplast is performed to form a new reconstructed egg cell. Following development of cloned embryos directly depends on successful association of the host and donor cells, and viability of the resulting hybrid (10-14).Electrofusion is commonly used technique for karyoplast-cytoplast fusion. A pulsed electric field applied to the cell membrane causes in it a reversible loss of integrity -transmembrane pores, after which the membrane becomes compressed and discharged. This makes possible the fusion of two contacting membranes, whose contact points open cytoplasmic channels for exchange of cytoplasmic colloid. As a result,...

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Improvement of canine somatic cell nuclear transfer procedure

Cited by 26 publications

References 35 publications

Generation of red fluorescent protein transgenic dogs

Generation of red fluorescent protein transgenic dogs

Microenvironnement tubaire: rôle dans la maturation des ovocytes canins in vivo et in vitro

Optimization of Parameters of the Oocyte Enucleation and Fusion With the Somatic Cell During Production of Mammalian Cloned Embryos

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