발생공학 국제심포지엄 및 학술대회 The 13th International Symposium on Developmental Biotechnology::한국동물생명공학회(구 한국동물번식학회)

1

Transchromosomic Cattle for Fully Human Antibody Production

Zhongde Wang

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.3

Therapeutic human polyclonal antibodies (hpAbs) produced from pooled plasma from human donors have been used in the treatment of a variety of human diseases. However, supply shortages and other restrictions such as donor-patient disease transmission concerns restrict the therapeutic use of plasma-derived hpAbs. Therefore, innovations are needed to improve both supply and usability of this promising biologic. We engineered transchromosomic (Tc) cattle carrying a human artificial chromosome (HAC) comprising the entire unrearranged human immunoglobulin (Ig) heavy-chain (hIGH), kappa-chain (hIGK), and lambdachain (hIGL) germline loci. To improve fully hpAbs production in the Tc cattle, we also sequentially knocked out the bovine Ig mu heavy-chains, bIGHM and bIGHML1, and the lambda gene cluster resulting in the production of triple knockout (TKO) Tc cattle. In this presentation, I will describe the construction of the HAC and the sequential gene targeting work to knock out the three bovine Ig loci. In addition, B cell development and the high level production of fully hpAbs in these Tc cattle will be presented.

2

Generation of Transgenic Pigs for Production of Humanized Antibody

KeunJung Kim, Jihye Lee, Kyungbon Lee, Jeongwoong Lee, MinKyu Kim

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.4

3

Ovum Pick-Up derived Embryo Production and Its Application

Il-Keun Kong, Jong-In Jin, NasserGhanem, Byung-Hyun Choi, Hyun-Tae Jo, Seong-Su Kim, Jae-Il Bang, A-Na Ha, Md Fakruzzaman, Han-Seul Park, Yu-Gon Kim, Pu-Rum Park, Suk-Hwan Song, Kyu-Woan Cho, Jung-Gyu Lee

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.5

4

Germ Cell Development in Non-Rodent Mammals

Ramiro Alberio

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.6

Primordial germ cells (PGCs) are the embryonic precursors of mature gametes. In mice, primordial germ cells originate from lineage restricted precursors that are segregated from the undifferentiated epiblast. During the development of germ cells a remarkable epigenetic reprogramming takes place, which is thought to be essential for preventing the inheritance of epimutations in the offspring. Furthermore, PGC can be isolated and will convert into naïve pluripotent embryonic germ cells (EG) when cultured in vitro. These cells have the capacity to differentiate into all somatic tissues and generate germline chimeras efficiently. These features suggest that PGC could be an alternative source of pluripotent cells in large animals species, where establishment of pluripotent cell lines from preimplantation embryos has been very challenging. Our lab investigates the kinetics of germ cell development in the pig embryo and studies the signalling pathways involved in the generation of germ cell precursors from pluripotent cells. We find that pig PGCs undergo epigenetic reprogramming over an extended period of time during the migratory phase, involving the erasure of H3K9-me2 and acquisition of H3K27me3. This is also followed by extensive DNA demethylation and erasure of imprinted loci in cells arriving to the gonadal primordium. We have also established that BMP4 plays an important in inducing the germline in this species, as shown in human and mouse species. Having established the critical timepoints of germ cell establishment we are now developing protocols for the establishment of EG cell lines as well as working on the differentiation of pluripotent cells into in vitro gametes. A detailed study of the kinetics of germ cell ontogeny in the pig will inform into the conserved mechanisms of germ cell development in mammals and is of critical importance for developing safe technologies that will enable the generation of EG cell lines and in vitro gametes.

5

Epigenetic Regulation of Germ Cell Development

Kwon-Ho Hong

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.7

6

New Insights into Mitochondrial Inheritance in Mice

Qing-Yuan Sun

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.8

7

Adipogenesis in Skeletal Muscle

Keitaro Yamanouchi

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.9

8

Production of Fat-1 Transgenic Dairy and Beef Cattle

Guang-Peng Li

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.10

As a more convinced technique, somatic cell nuclear transfer (SCNT) has been successfully applied in the production of transgenic farm animals although the cloning efficiency is still low. One of the significant advantages of transgenesis is in rapid genetic improvement of interested traits such as growth rate, muscle growth, disease resistance. In this talk, I focused on the improved modifications of bovine SCNT protocols and its application in the production of fat-1 transgenic dairy and beef cattle. We generated mfat-1 transgenic cattle expressed a Caenorhabditis elegans gene, mfat-1, encoding an n-3 fatty acid desaturase. Fatty acids analysis of tissue and milk derived from transgenic positive cow showed that all of the examined n-3 polyunsaturated fatty acids (PUAFs) were greatly increased and simultaneously the n-6 PUAFs decreased. A significantly reduction of n-6/n-3 ratios (p<0.05) in both tissue and milk were observed. Similar results were obtained in the transgenic beef cattle. Thus, the results indicate that fat-1 transgenic cattle can produce meat and milk enriched in n-3 fatty acids, which probably become an efficient and economical approach to meet the increasing demand for n-3 PUAFs.

9

Generation of Cloned Embryos from Porcine Induced Pluripotent Stem Cells at Metaphase

Zhong Zheng, Yong-Xun Jin, Chang-Kyu Lee, Nam-Hyung Kim, Sang Hwan Hyun

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.11

10

Production of Elite Military Sniffing Dogs by Somatic Cell Nuclear Transfer

Dong-Hoon Kim, Mi-Kyung Choi, Jin-Gu No, Jong-Ju Park, Dong-Hyeon Yeom, Dong-Kyo Kim, Bong-Hwan Choi, Jae Gyu Yoo, Byoung-Chul Yang, Jin-Ki Park

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.17

11

Comparison of Fresh with Vitrified White-colored Hanwoo Cloned Embryos

Tae-Suk Kim, Sang-Ki Baek, Song-Yi Moon, Sang-Jin Jin, Yeoung-Gyu Ko, Sung-Woo Kim, Hae-Geum Park, Hwan-Hoo Seong, Joon-Hee Lee

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 pp.18-19

12

Knockout Beta-Lactoglobulin (BLG) in Bovine Embryos using RNA-guided Cas9 System

Woo Jae Choi, Choong Il Lee, Ji Hyun Lee, Soo Young Yum, Byeong Chun Lee, Jin Soo Kim, Goo Jang

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.20

13

Establishment of hEPO Cloned Embryo Production System in Bovine

A-Na Ha, Su-Jin Cho, Sang-Ryeul Lee, Han-Seul Park, Jong-In Jin, Kyeong-Lim Lee, Dae-Hwan Ko, Sang-June Eum, Do-Joong Yoon, Jae-Il Bang, Zhongde Wang, Il-Keun Kong

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.21

14

Improved Development Competence using Immortalized Cells as Donor in Bovine Nuclear Transfer

Sisitha Ramachandra, Don Buddika Oshadi Malaweera, Junbo Wu, Seungkyu Oh, Seunghwan Kim, Woojae Choi, Goo Jang, Jongki Cho

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.22

15

Antioxidant Treatment during Somatic Cell Nuclear Transfer Procedures Prevents the Mitochondria and DNA Damages and Enhances the Nuclear Reprogramming of Cloned Bovine Embryos

Hyo-Kyung Bae, In-Sun Hwang, Choon-keun Park, Boo-Keun Yang, Hee-Tae Cheong

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.23

16

Spermatogonial Stem Cells from the Testicle of New-Born Piglets can support Somatic Cell Nucleus Transferred Embryo Development In Vitro

Joohyeong Lee, Yongjin Lee, Bola Park, Fazle Elahi, Min Hee Park, Jung Hoon Choi, Seung Tae Lee, Eunsong Lee

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.24

17

Somatic Cell Nuclear Transfer using Vitrified-Thawed Bovine MII Oocytes

Min-Jee Park, Eun-Young Kim, Seung-Eun Lee, Hyun-Yong Choi, Jeremiah Ji-Man Moon, Se-Pill Park

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.25

18

Comparative Sequence Analysis of Porcine OCT4 Promoter Region for Developing Reprogramming Reporter System

Jae Yeon Hwang, Dong-Chan Son, Kwang-hwan Choi, Chang-kyu Lee

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 pp.26-27

19

Post-Death Cloning of Endangered Jeju Black Cattle (Korean Native Cattle): Fertility and Serum Chemistry in a Cloned Bull and Cow and Their Offspring

Dong-Hwan Song, Eun-Young Kim, Min-Jee Park, Seung-Eun Lee, Hyun-Yong Choi, Jeremiah Ji-Man Moon, Chang Eon Oh, Young Hoon Kim, Seong Ho Mun, Moon Suck Ko, Se-Pill Park

한국동물생명공학회(구 한국동물번식학회) 발생공학 국제심포지엄 및 학술대회 Vision of Animal Reproductive Biotechnology ; from Basic Research to Practical Applications 2013.10 p.28