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Production of Cloned Jeju Black Cattle (Korean Cattle) from SCNT Embryo using Vitrification, One-Step Dilution and Direct Transfer Technique  

Kim, Eun-Young (Mirae Biotech)
Park, Min-Jee (Mirae Biotech)
Kim, Jae-Youn (Mirae Biotech)
Park, Hyo-Young (Mirae Biotech)
Noh, Eun-Ji (Mirae Biotech)
Noh, Eun-Hyung (Mirae Biotech)
Song, Dong-Hwan (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, Institue for Livestock Promotion)
Mun, Seong-Ho (Jeju National University Stem Cell Research Center)
Lee, Dong-Sun (Jeju National University)
Ko, Moon-Suck (National Institute of Subtropical Agriculture)
Riu, Key-Zung (Jeju National University)
Park, Se-Pill (Mirae Biotech)
Publication Information
Reproductive and Developmental Biology / v.35, no.1, 2011 , pp. 77-83 More about this Journal
Abstract
One-step dilution and direct transfer would be a practical technique for the field application of frozen embryo. This study was to examine whether Jeju Black Cattle (JBC, Korean Cattle) can be successfully cloned from vitrified and one-tep diluted somatic cell nuclear transfer (SCNT) blastocyst after direct transfer. For vitrification, JBC-SCNT blastocysts were serially exposed in glycerol (G) and ethylene glycol (EG) mixtures [10%, (v/v) G for 5 min., 10% G plus 20% EG (v/v) for 5 min., and 25% G plus 25% EG (v/v) for 30 sec.] which is diluted in 10% FBS added D-PBS. And then SCNT blastocysts were loaded in 0.25 ml mini straw, placed in cold nitrogen vapor for 3 min. and then plunged into $LN_2$. One-step dilution in straw was done in $25^{\circ}C$ water for 1 min, by placing vertically in the state of plugged-end up and down for 0.5 min, respectively. When in vitro developmental capacity of vitrified SCNT blastocyst was examined at 48 h after one-step dilution, hatched rate (56.4%) was slightly lower than that of control group (62.5%). In field trial, when the vitrified-thawed SCNT blastocysts were transferred into uterus of synchronized 5 recipients, a cloned female JBC was delivered by natural birth on day 299 and healthy at present. In addition, when the short tandem repeat marker analysis of the cloned JBC was evaluated, microsatellite loci of 11 numbers was perfectly matched genotype with donor cell (BK94-14). This study suggested that our developed vitrification and one-step dilution technique can be applied effectively on field trial for cloned animal production, which is even no longer in existence.
Keywords
Jeju Black Cattle; SCNT blastocyst; Vitrification; One-step dilution; Direct transfer;
Citations & Related Records
Times Cited By KSCI : 2  (Citation Analysis)
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