Improving porcine SCNT efficiency by selecting donor cells size

Deling Jiao; Wenmin Cheng; Xiaolin Zhang; Yifan Zhang; Jianxiong Guo; Zhuo Li; Dejia Shi; Zhe Xiong; Yubo Qing; Muhammad Ameen Jamal; Kaixiang Xu; Hong-Ye Zhao; Hong-Jiang Wei

doi:10.1080/15384101.2021.1980983

Improving porcine SCNT efficiency by selecting donor cells size

Cell Cycle. 2021 Nov;20(21):2264-2277. doi: 10.1080/15384101.2021.1980983. Epub 2021 Sep 29.

Authors

Deling Jiao^{1

2

3}, Wenmin Cheng^{1

2

3}, Xiaolin Zhang^{1

2

3}, Yifan Zhang^{1

2

3}, Jianxiong Guo^{1

2

3}, Zhuo Li^{1

2

3}, Dejia Shi^{1

2

3}, Zhe Xiong^{1

2

3}, Yubo Qing^{1

2

3

4}, Muhammad Ameen Jamal^{1

2

3}, Kaixiang Xu^{1

2

3}, Hong-Ye Zhao^{1

2

5}, Hong-Jiang Wei^{1

2

4

5}

Affiliations

¹ Key laboratory for porcine gene editing and xenotransplantation in Yunnan Province, Yunnan Agricultural University, Kunming, China.
² Xenotransplantation Research Engineering Center in Yunnan Province, Yunnan Agricultural University, Kunming, China.
³ College of Animal Science and Technology, Yunnan Agricultural University, Kunming, China.
⁴ College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China.
⁵ State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming, China.

Abstract

Considerable advancements have recently been achieved in porcine somatic cell nuclear transfer (SCNT), but the efficiency remains low. Donor cell size might play an important role in SCNT, but its effects in pigs remain unclear. This study aimed to evaluate the efficiency of porcine SCNT by selecting donor cells of suitable size. Porcine fetal fibroblasts (PFFs) were divided into three groups, group S (small, d ≤ 13 μm), group M (medium, 13 μm<d ≤ 18 μm), and group L (large, d > 18 μm), and their biological characteristics were analyzed. Next, SCNT was performed using PFFs of different sizes to evaluate the developmental potential of reconstructed embryos. The data showed that PFFs in groups S, M and L accounted for 17.5%, 47.7% and 34.8% of cells, respectively. Morphologically, cells in group S exhibited clear and regular cell membranes and nuclei, whereas cells in groups M and L displayed varying degrees of cell membrane protuberance, karyo-pyknosis, autophagy and mitochondrial abnormalities. In addition, the growth status and proliferation capabilities of cells in group S were significantly better than those of group M and group L. The percentage of cells at G0/G1 in group S and M were significantly greater than group L. The senescence rate of group S was lower than group M and group L. The apoptosis rate of group S was significantly lower than that of group L but comparable to that of group M . The cleavage rate of group S was also significantly greater than that of group M but comparable to that of group L . The blastocyst rate of group S was significantly greater than that of group M and group L. The blastocyst cell numbers of group S were also significantly greater than those of group M and group L. These findings suggested that small PFFs with a diameter of less than 13 μm are more suitable donor cells for SCNT in pigs.Abbreviations: DMEM: Dulbecco's modified Eagle's medium; FBS: Fetal bovine serum; PBS: Phosphate buffer saline; PFFs: Porcine fetal fibroblast cells; SCNT: Somatic cell nuclear transfer.

Keywords: Pig; SCNT; cell size; reconstructed embryo development.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Blastocyst* / metabolism
Cell Size
Cloning, Organism
Embryo, Mammalian
Embryonic Development
Female
Fetus
Fibroblasts / metabolism
Nuclear Transfer Techniques*
Pregnancy
Swine

Grants and funding

This work was supported by grants from National Key R&D Program of China [Grant No. 2019YFA0110700], Major Science and Technology Project of Yunnan Province [Grant No. 202102AA310047] and Yunling Scholars Program of Yunnan Province .