The effect of electrical field strength on activation and development of cloned caprine embryos

Shen, Perng-Chih; Lee, S.N.; Wu, J.S.; Huang, Jeng-Fang; Chu, F.H.; Chang, C.C.; Kung, Jung-Chang; Lin, Hsiu-Lien; Chen, L.R.; Shiau, Jen-Wen; Yen, N. T.; Cheng, W. T. K.

doi:10.1016/j.anireprosci.2005.05.025

Cited by 18 publications

(9 citation statements)

References 43 publications

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“…Goats, being an important source of milk, meat, hair, and manure, is spread throughout the world and helps in sustaining the economy of the b AU2 farmers. In the past, several attempts have been made to produce cloned (Lan et al, 2006;Shen et al, 2006) and transgenic goats (Baguisi et al, 1999;Keefer et al, 2001) using the micromanipulator system. However, no information is available to date, on the application of a HMC technique in goats, except a study by Akshey et al (2008).…”

Section: Introductionmentioning

confidence: 99%

Hand-Made Cloned Goat (Capra hircus) Embryos—A Comparison of Different Donor Cells and Culture Systems

Akshey

Malakar²,

De³

et al. 2010

Cellular Reprogramming

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Nuclear transfer is a very effective method for propagation of valuable, extinct, and endangered animals. Hand-made cloning (HMC) is an efficient alternative to the conventional micromanipulator-based technique in some domestic species. The present study was carried out for the selection of suitable somatic cells as a nuclear donor and development of an optimum culture system for in vitro culture of zona-free goat cloned embryos. Cleavage and blastocyst rates were observed 72.06 ± 2.94% and 0% for fresh cumulus cells, 81.95 ± 3.40% and 12.74 ± 2.12% for cultured cumulus cells, and 92.94 ± 0.91% and 23.78 ± 3.33% for fetal fibroblast cells, respectively. There was a significant (p < 0.05) increase in blastocyst production in goats when cultured on a flat surface (FS) (23.78 ± 3.33 %) than well of wells (WOW) (15.84 ± 2.12 %) and microdrops (MD) (0.7 ± 0.7%). Furthermore, cleavage and blastocyst production rates were significantly (p < 0.05) more in the WOW (15.84 ± 2.12%) than the MD (0.7 ± 0.7%) system. The quality of HMC blastocysts was studied by differential staining. Genetic similarity was confirmed by polymerase chain reaction (PCR)-based amplification of the second exon of the MHC class II DRB gene, which gave similar bands in electrophoresis (286 bp) both in cloned embryos and donor cells. In conclusion, the present study describes that the fetal fibroblast cell is a suitable candidate as nuclear donor, and the flat surface culture system is suitable for zona-free blastocyst development by the hand-made cloning technique in the goat.

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Section: Introductionmentioning

confidence: 99%

Hand-Made Cloned Goat (Capra hircus) Embryos—A Comparison of Different Donor Cells and Culture Systems

Akshey

Malakar²,

De³

et al. 2010

Cellular Reprogramming

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“…After transfer, the karyoplast-ooplasm couplets were washed in fresh medium to remove CB for at least 15 min prior to electrofusion with an Electro-Cell Manipulator (BTX200, San Diego, CA) in Ca 2þ -free fusion solution (0.3 M mannitol, 0.1 mM MgCL 2 6H 2 O and 0.5 mM HEPES). Membrane fusion was induced by applying a DC pulse (1.7 kV=cm, 15 msec) according to a protocol described by Shen et al (2006Shen et al ( , 2008. After fusion, the reconstructed oocytes including HSsp-Coop, Csp-HSoop, and Csp-Coop groups were loaded with fura-2 AM as described previously.…”

mentioning

confidence: 99%

Calcium Release and Development of Heat-Shocked Porcine Oocytes after Nucleus-Ooplasm Reconstruction

Tseng

Liu

Lin

et al. 2009

Cloning and Stem Cells

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We determined the effect of heat shock (HS) on the alterations of development and calcium releasing capacity of nuclear-ooplasmic reconstructed porcine oocytes stimulated by thimerosal. The non-HS (39 degrees C) and the HS2h (41.5 degrees C for 2 h) matured oocytes were enucleated and their spindles/chromosomes were exchanged between these two groups followed by parthenogenetic activation. In the Control group (Csp-Coop), the non-HS spindle (Csp) was transferred to the non-HS ooplasm (Coop). Blastocyst and cleavage rates were higher in both Csp-HSoop (non-HS spindle transferred to the HS ooplasm) and HSsp-Coop (HS spindle transferred to non-HS ooplasm) reconstructed oocytes, but no difference was detected in the average cell number per blastocyst. However, intracellular calcium concentrations ([Ca(2+)](i)) generally declined (p < 0.05) in the reconstructed HS oocytes, with a greater blastocyst rate after parthenogenetic activation. In the present study, time for the completion of spindle transfer in these oocytes was 1-2 h, during which some physiological remodeling or adaptation might have been occurred in the oocytes. Therefore, changes in heat-shock protein70 (HSP70) expression and developmental competence of the HS2h oocytes with 1 or 2 h of recovery time under normal culture temperature (39 degrees C) were examined. The results showed that the expression of HSP70 in the HS2h oocytes was higher (p < 0.05) than those had recovery incubation for 1 h (HC1h) after HS, but the cleavage and blastocyst rates were greater (p < 0.05) in the HC1h group. We demonstrated that a recovery period prior to activation of porcine oocytes and reconstructed oocytes is beneficial to further development. Heat shock to either the karyoplast or the ooplasm enhances embryonic development but reduces intracellular calcium release in the cloned porcine oocytes.

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“…Since 1997 when the first SCNT sheep was announced (Wilmut et al, 1997), offspring have been produced in a number of different species (Vajta and Gjerris, 2006). The cloning of goats by SCNT has also been reported (Baguisi et al, 1999;Keefer et al, 2001Keefer et al, , 2002Reggio et al, 2001;Zou et al, 2001Zou et al, , 2002Behboodi et al, 2004;Chen et al, 2006;Lan et al, 2006;Shen et al, 2006).…”

mentioning

confidence: 98%

“…According to the previous report (Shen et al, 2006), the chemical fusion solution consisted of 0.32M mannitol, 100µM CaCl 2 , 100µM MgSO 4 , 0.5mM Hepes, and 0.01 mg/ml BSA. According to our previously established method , electrofusion and activation were undertaken after chemical fusion.…”

Section: Somatic Cell Nuclear Transfer Protocolmentioning

confidence: 99%

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Production of Boer goat (Capra hircus) by nuclear transfer of cultured and cryopreserved fibroblast cells into slaughterhouse-derived oocytes

Tao¹,

Han²,

Zhang³

et al. 2009

CZECH J ANIM SCI

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ABstrAct:The present study reports a birth of goat clone produced by nuclear transfer of cultured and cryopreserved fibroblast cells into slaughterhouse-derived oocytes. The donors of somatic cells were fibroblasts derived from the ear skin of a Boer goat while the recipient ooplasm was in vitro matured oocytes of Huanghuai white goat, an Anhui native goat species. The reconstructed embryos were cultured in vitro and then the morphologically normal embryos were transferred to the surrogates. The reconstructed embryos were surgically transferred into 37 recipient surrogates, Huanghuai white goats with natural oestrus. Five of them were treated with hCG after transfer. Among them, one was pregnant and gave birth to a live kid. Due to the improper delivery aid, the cloned kid died accidentally after birth. The cloned kid was then anatomised to observe the viscera development, and the results showed that the organs were normal. Paraffin tissue slices were prepared and stained to ensure the viscera development further, and the results suggested that the organs also developed well in spite of incipient hydropericardium. The microsatellite analysis identified the cloning. It is suggested that the optimised nuclear transfer protocol and proper hCG treatment lead to the successful birth of a goat clone.Keywords: goat; fibroblast; clone; hCG 449Czech J. Anim. Sci., 54, 2009 (10): 448-460 Original Paper are needed to elucidate to what extent the origin of the oocyte affects the development of resultant reconstructed embryos into foetuses and possibly offspring in the goat. The administration of exogenous hormones like human chorionic gonadotropin (hCG) was used in cattle, sheep and goat (Fonseca and Torres, 2005). This hormonal administration can efficiently promote the ovulation of the first-wave dominant follicle forming the accessory corpus luteum.The Boer goat, a famous subspecies originated from South Africa, has advantages primarily for meat production. In addition to its docility and adaptability to various environments and feeds, this breed grows faster and its meat is popular among consumers. Moreover, the Boer goat phenotype including a brown and white head with big ears may be utilised as a marker in genetic studies.The present study was therefore undertaken to initiate Boer goat cloning by SCNT using native goats as oocyte donors and foster mothers to examine whether the hCG injection to recipients after embryo transfer improves the success rate. MAtErIAL AND MEtHODs selection of animalsAn adult Boer goat was used as a donor of somatic cells. Huanghuai white goat, an Anhui native goat species, was used as donors of oocytes as well as recipients of SCNT embryos. More than 70 goats were raised on a farm as recipient candidates, receiving general health care, and their oestrus was also observed. Exclusively healthy individuals were selected for the present experiments. Thirty-seven recipient surrogates were used altogether in the present study.Blood samples were obtained by jugular venipuncture using heparin-vac...

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The effect of electrical field strength on activation and development of cloned caprine embryos

Cited by 18 publications

References 43 publications

Hand-Made Cloned Goat (Capra hircus) Embryos—A Comparison of Different Donor Cells and Culture Systems

Hand-Made Cloned Goat (Capra hircus) Embryos—A Comparison of Different Donor Cells and Culture Systems

Calcium Release and Development of Heat-Shocked Porcine Oocytes after Nucleus-Ooplasm Reconstruction

Production of Boer goat (Capra hircus) by nuclear transfer of cultured and cryopreserved fibroblast cells into slaughterhouse-derived oocytes

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