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O型血韩国本土猪细胞系的建立、鉴定及其通过体细胞核移植支持胚胎发育的效率

Establishment and identification of cell lines from type O blood Korean native pigs and their efficiency in supporting embryonic development via somatic cell nuclear transfer.

作者信息

Taweechaipaisankul Anukul, Kim Geon A, Jin Jun-Xue, Yeom Su Cheong, Lee Byeong Chun

机构信息

Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul 08826, Korea.

Institutes of Green Bio Science and Technology, Seoul National University, Pyeongchang 25354, Korea.

出版信息

J Vet Sci. 2018 Jul 31;19(4):492-499. doi: 10.4142/jvs.2018.19.4.492.

Abstract

Due to their similarities with humans in anatomy, physiology, and genetics miniature pigs are becoming an attractive model for biomedical research. We aim to establish and evaluate blood type O cells derived from Korean native pig (KNP), a typical miniature pig breed in Korea. Ten cell lines derived from 8 KNP piglets and one adult female KNP (kidney and ear tissues) were established. To confirm the presence of blood type O, genomic DNA, fucosyltransferase (FUT) expression, and immunofluorescence staining were examined. Additionally, fluorescence-activated cell sorting and somatic cell nuclear transfer were performed to investigate the normality of the cell lines and to evaluate their effectiveness in embryo development. We found no significant bands corresponding to specific blood group A, and no increase in FUT expression in cell lines derived from piglets No. 1, No. 4, No. 5, No. 8, and the adult female KNP; moreover, they showed normal levels of expression of α 1,3-galactosyltransferase and cytidine monophosphate-N-acetylneuraminic acid hydroxylase. There was no significant difference in embryo development between skin and kidney fibroblasts derived from the blood type O KNPs. In conclusion, we successfully established blood type O KNP cell lines, which may serve as a useful model in xenotransplantation research.

摘要

由于小型猪在解剖学、生理学和遗传学方面与人类相似,它们正成为生物医学研究中一种有吸引力的模型。我们旨在建立并评估源自韩国本土猪(KNP,韩国一种典型的小型猪品种)的O型血细胞系。从8只KNP仔猪和1只成年雌性KNP(肾脏和耳部组织)中建立了10个细胞系。为了确认O型血的存在,检测了基因组DNA、岩藻糖基转移酶(FUT)表达和免疫荧光染色。此外,进行了荧光激活细胞分选和体细胞核移植,以研究细胞系的正常性并评估它们在胚胎发育中的有效性。我们发现没有与特定A型血对应的明显条带,并且在来自第1、4、5、8号仔猪和成年雌性KNP的细胞系中FUT表达没有增加;此外,它们显示出α1,3-半乳糖基转移酶和胞苷单磷酸-N-乙酰神经氨酸羟化酶的正常表达水平。源自O型血KNP的皮肤和肾脏成纤维细胞在胚胎发育方面没有显著差异。总之,我们成功建立了O型血KNP细胞系,这可能在异种移植研究中作为一个有用的模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d3bb/6070591/227c488da22b/jvs-19-492-g001.jpg

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