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In Vitro Development of Somatic Cell Nuclear Transfer Embryo Treated with Flavonoid and Production of Cloned Jeju Black Cattle  

Kim, Eun-Young (Mirae Biotech)
Kim, Yeon-Ok (Mirae Biotech)
Kim, Jae-Youn (Mirae Biotech)
Park, Min-Jee (Mirae Biotech)
Park, Hyo-Young (Mirae Biotech)
Han, Young-Joon (Jeju National University Stem Cell Research Center)
Mun, Seong-Ho (Jeju National University Stem Cell Research Center)
Oh, Chang-Eon (Jeju National University Stem Cell Research Center)
Kim, Young-Hoon (Jeju Special Self-Governing Province, Institute for Livestock Promotion)
Lee, Sung-Soo (National Institute of Subtropical Agriculture)
Ko, Moon-Suck (National Institute of Subtropical Agriculture)
Park, Se-Pill (Mirae Biotech)
Publication Information
Reproductive and Developmental Biology / v.34, no.3, 2010 , pp. 127-134 More about this Journal
Abstract
This study was to investigate the effect of flavonoid treatment on in vitro development of bovine somatic cell nuclear transfer (SCNT) embryos, and their pregnancy and delivery rate after embryo transfer into recipient. In experiment 1, to optimize the flavonoid concentration, parthenogenetic day 2 ($\geq$ 2-cell) embryos were cultured in 0 (control), 1, 10 and $20\;{\mu}M$ flavonoid for 6 days. In the results, in vitro development rate was the highest in $10\;{\mu}M$ flavonoid group (57.1%) among treatment groups (control, 49.5%; $1\;{\mu}M$, 54.2%; $20\;{\mu}M$, 37.5%), and numbers of total and ICM cells were significantly (p<0.05) higher in $10\;{\mu}M$ flavonoid group than other groups. We found that $10\;{\mu}M$ flavonoid treatment can significantly (p<0.05) decrease the apoptotic index and derive high expression of anti-oxidant, anti-apoptotic, cell growth and development marker genes such as Mn-SOD, Survivin, Bax inhibitor, Glut-5, In-tau, compared to control group. In experiment 2, to produce the cloned Jeju Black Cattle, beef quality index grade 1 bull somatic cells were transferred into enucleated bovine MII oocytes and reconstructed embryos were cultured in $10\;{\mu}M$ flavonoid added medium. When the in vitro produced day 7 or 8 SCNT blastocysts were transferred into a number of recipients, $10\;{\mu}M$ flavonoid treatment group presented higher pregnancy rate (10.2%, 6/59) than control group (5.9%, 2/34). Total three cloned Jeju Black calves were born. Also, two cloned calves in $10\;{\mu}M$ flavonoid group were born and both were all healthy at present, while the one cloned calf born in control group was dead one month after birth. In addition, when the result of short tandem repeat marker analysis of each cloned calf was investigated, microsatellite loci of 11 numbers matched genotype between donor cell and cloned calf tissue. These results demonstrated that the flavonoid addition in culture medium may have beneficial effects on in vitro and in vivo developmental capacity of SCNT embryos and pregnancy rate.
Keywords
Bovine; SCNT; Flavonoid; Jeju Black Cattle; Pregnancy;
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Times Cited By KSCI : 2  (Citation Analysis)
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