title: Production of Pigs by Hand-Made Cloning Using Mesenchymal Stem Cells and Fibroblasts.
abstract: Mesenchymal stem cells (MSCs) exhibited self-renewal and less differentiation, making the MSCs promising candidates for adult somatic cell nuclear transfer (SCNT). In this article, we tried to produce genome identical pigs through hand-made cloning (HMC), with MSCs and adult skin fibroblasts as donor cells. MSCs were derived from either adipose tissue or peripheral blood (aMSCs and bMSCs, respectively). MSCs usually showed the expression pattern of CD29, CD73, CD90, and CD105 together with lack of expression of the hematopoietic markers CD34and CD45. Flow cytometry results demonstrated high expression of CD29 and CD90 in both MSC lines, while CD73, CD34, and CD45 expression were not detected. In contrary, in reverse transcription-polymerase chain reaction (RT-PCR) analysis, CD73 and CD34 were detected indicating that human antibodies CD73 and CD34 were not suitable to identify porcine cell surface markers and porcine MSC cellular surface markers of CD34 might be different from other species. MSCs also had potential to differentiate successfully into chondrocytes, osteoblasts, and adipocytes. After HMC, embryos reconstructed with aMSCs had higher blastocyst rate on day 5 and 6 than those reconstructed with bMSCs and fibroblasts (29.6% ± 1.3% and 41.1% ± 1.4% for aMSCs vs. 23.9% ± 1.2% and 35.5% ± 1.6% for bMSCs and 22.1% ± 0.9% and 33.3% ± 1.1% for fibroblasts, respectively). Live birth rate per transferred blastocyst achieved with bMSCs (1.59%) was the highest among the three groups. This article was the first report to compare the efficiency among bMSCs, aMSCs, and fibroblasts for boar cloning, which offered a realistic perspective to use the HMC technology for commercial breeding.
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .|2 BGI-Shenzhen , Shenzhen, China .
authors.forename: Zhenzhen
authors.initials: Z
authors.lastname: Yang
authors.affiliation: 2 BGI-Shenzhen , Shenzhen, China .|3 Central Queensland University , Rockhampton, Australia .
authors.forename: Gábor
authors.initials: G
authors.lastname: Vajta
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .|2 BGI-Shenzhen , Shenzhen, China .
authors.forename: Ying
authors.initials: Y
authors.lastname: Xu
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .|2 BGI-Shenzhen , Shenzhen, China .
authors.forename: Jing
authors.initials: J
authors.lastname: Luan
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .|2 BGI-Shenzhen , Shenzhen, China .
authors.forename: Mufei
authors.initials: M
authors.lastname: Lin
authors.affiliation: 2 BGI-Shenzhen , Shenzhen, China .
authors.forename: Cong
authors.initials: C
authors.lastname: Liu
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .
authors.forename: Jianing
authors.initials: J
authors.lastname: Tian
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .
authors.forename: Hongwei
authors.initials: H
authors.lastname: Dou
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .|2 BGI-Shenzhen , Shenzhen, China .
authors.forename: Yong
authors.initials: Y
authors.lastname: Li
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .
authors.forename: Tianbin
authors.initials: T
authors.lastname: Liu
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .
authors.forename: Yijie
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .
authors.forename: Wenxian
authors.initials: W
authors.lastname: Yang
authors.affiliation: 2 BGI-Shenzhen , Shenzhen, China .|4 Department of Biomedicine, University of Aarhus , Aarhus C, Denmark .
authors.forename: Lars
authors.initials: L
authors.lastname: Bolund
authors.affiliation: 2 BGI-Shenzhen , Shenzhen, China .
authors.forename: Huanming
authors.initials: H
authors.lastname: Yang
authors.affiliation: 1 BGI Ark Biotechnology Co., LTD (BAB) , Shenzhen, China .|2 BGI-Shenzhen , Shenzhen, China .
authors.forename: Yutao
authors.initials: Y
authors.lastname: Du
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology* / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D013552:Swine