五指山小型猪近交系异种移植产业化研发进展

冯书堂; 戴一凡; 章金刚; 潘志强

doi:10.3969/j.issn.1674-7445.2018.06.014

五指山小型猪近交系异种移植产业化研发进展

doi: 10.3969/j.issn.1674-7445.2018.06.014

100007 北京盖兰德生物科技有限公司（冯书堂）; 南京医科大学（戴一凡）; 军事科学院军事医学研究院卫生勤务与血液研究所（章金刚）; 北京同仁医院（潘志强）

基金项目:

国家重点研发计划重点专项 2017YFC11037000

国家重点研发计划重点专项 2017YFD0501606

详细信息

通讯作者:
冯书堂，男，1945年生，1975年至2015年于中国农业科学院北京畜牧兽医研究所从事畜牧科学研究，研究员，博士研究生导师，2016年至今于北京市盖兰德科技公司任名誉董事长、首席科学家，主要从事动物繁育技术和实验用小型猪近交系培育与产业化研发，Email：fst508@sina.com

中图分类号: R617, S828
计量
- 文章访问数: 194
- HTML全文浏览量: 124
- PDF下载量: 11
- 被引次数: 0
出版历程
- 收稿日期: 2018-08-16
- 网络出版日期: 2021-01-19
- 刊出日期: 2018-11-15

摘要

摘要: 为了早日实现五指山小型猪（WZSP）近交系异种移植产业化目标，努力推进近交繁育获得近交系，同时开展克服异种移植免疫排斥和猪内源性逆转录病毒（PERV）传染生物安全性两大难题的研发内容。本文从WZSP近交系双基因敲除克隆猪繁育成功、猪-猴异种角膜内皮移植取得突破性进展、WZSP近交系PERV无传染性群体建立、WZSP近交系是理想的动物模型和异种移植供体等方面，介绍WZSP近交系异种移植产业化的研发进展。
- 五指山小型猪 /
- 近交系猪 /
- 异种移植 /
- 免疫排斥反应 /
- α-1, 3-半乳糖基转移酶基因敲除（αGTKO） /
- 猪内源性逆转录病毒（PERV） /
- 非α-1, 3-半乳糖（Gal）抗原 /
- CRISPR/Cas9基因编辑技术

HTML全文

图 1 GGTA1/β₄GalNT2双基因敲除猪的生长发育情况

Figure 1. Growth and development of GGTA1/β₄GalNT2 double gene knockout pigs

下载: 全尺寸图片幻灯片

图 2 WZSP近交系-恒河猴板层角膜移植成功

图示不同观察时间板层角膜移植术后植片均保持透明

Figure 2. Lamellar corneal transplantation of WZSP inbred line-Rhesus Monkey

下载: 全尺寸图片幻灯片

图 3 初步筛选与确证实验的技术路线

HEK293细胞为人胚胎肾细胞; PBMC为外周血单核细胞

Figure 3. Technical route of preliminary screening and confirming experiment

下载: 全尺寸图片幻灯片

表 1 中国小型猪体内PERV的检测与筛查结果^[21]

Table 1. Detection and screening result of PERV in miniature pigs in China

品系	产地	n	PERV亚型感染（只）
品系	产地	n	A	B	C
版纳小型猪	云南省	16	8	15	5
贵州小型猪	贵州省	36	18	32	9
剑河香猪	贵州省	20	11	19	15
巴马小型猪	广西省	101	87	98	21
巴马香猪	广西省	12	12	12	9
巴马香猪	上海	28	6	27	19
巴马香猪	重庆	20	16	20	17
贵州小型猪	重庆	22	18	20	9
农大小型猪	北京	26	18	26	2
五指山小型猪	北京	67	65	63	0

下载: 导出CSV

参考文献(25)

[1]	DAI Y, VAUGHT TD, BOONE J, et al. Targeted disruption of the alpha1, 3-galactosyltransferase gene in cloned pigs[J]. Nat Biotechnol, 2002, 20(3): 251-255. doi: 10.1038/nbt0302-251
[2]	LAI L, KOLBER-SIMONDS D, PARK KW, et al. Production of alpha-1, 3-galactosyltransferase knockout pigs by nuclear transfer cloning[J]. Science, 2002, 295(5557): 1089-1092. doi: 10.1126/science.1068228
[3]	DUQUESNOY RJ, 李幼平.移植免疫生物学[M].北京:科学出版社, 2000: 617-618.
[4]	ESTRADA JL, MARTENS G, LI P, et al. Evaluation of human and non-human primate antibody binding to pig cells lacking GGTA1/CMAH/β4GalNT2 genes[J]. Xenotransplantation, 2015, 22(3): 194-202. DOI: 10.1111/xen.12161.
[5]	HIGGINBOTHAM L, MATHEWS D, BREEDEN CA, et al. Pre-transplant antibody screening and anti-CD154 costimulation blockade promote long-term xenograft survival in a pig-to-primate kidney transplant model[J]. Xenotransplantation, 2015, 22(3): 221-230. DOI: 10.1111/xen.12166.
[6]	MOHIUDDIN MM, SINGH AK, CORCORAN PC, et al. Chimeric 2C10R4 anti-CD40 antibody therapy is critical for long-term survival of GTKO.hCD46.hTBM pig-to-primate cardiac xenograft[J]. Nat Commun, 2016, 7: 11138. DOI: 10.1038/ncomms11138.
[7]	VAN DER WINDT DJ, BOTTINO R, CASU A, et al. Long-term controlled normoglycemia in diabetic non-human primates after transplantation with hCD46 transgenic porcine islets[J]. Am J Transplant, 2009, 9(12): 2716-2726. DOI: 10.1111/j.1600-6143.2009.02850.x.
[8]	SHAH JA, NAVARRO-ALVAREZ N, DEFAZIO M, et al. A bridge to somewhere: 25-day survival after pig-to-baboon liver xenotransplantation[J]. Ann Surg, 2016, 263(6): 1069-1071. DOI: 10.1097/SLA.0000000000001659.
[9]	LAIRD C, BURDORF L, PIERSON RN 3RD. Lung xenotransplantation: a review[J]. Curr Opin Organ Transplant, 2016, 21(3): 272-278. DOI: 10.1097/MOT.0000000000000311.
[10]	YANG L, GÜELL M, NIU D, et al. Genome-wide inactivation of porcine endogenous retroviruses (PERVs)[J]. Science, 2015, 350(6264): 1101-1104. DOI: 10.1126/science.aad1191.
[11]	NIU D, WEI HJ, LIN L, et al. Inactivation of porcine endogenous retrovirus in pigs using CRISPR-Cas9[J]. Science, 2017, 357(6357): 1303-1307. DOI: 10.1126/science.aan4187.
[12]	FANG X, MOU Y, HUANG Z, et al. The sequence and analysis of a Chinese pig genome[J]. Gigascience, 2012, 1(1):16. DOI: 10.1186/2047-217X-1-16.
[13]	MU YL, LIU L, FENG ST, et al. Identification of the miniature pig inbred line by skin allograft[J]. J Integr Agricul, 2015, 14(7): 1376-1382. DOI: 10.1016/S2095-3119(14)60976-X.
[14]	FENG ST, ZHANG XL, WANG TP. The progress on cultivation and identification of the first Wuzhishan inbred mini-pig in China[J]. Agri Res Tech: Open Access J, 2017, 12(4): 555858. DOI: 10.19080/ARTOAJ.2017.12.555858.
[15]	冯书堂, 高倩, 刘岚.哺乳动物近交系资源创新百年[J].遗传, 2016, 38(3): 181-195. DOI: 10.16288/j.yczz.15-430. FENG ST, GAO Q, LIU L. Resources of mammalian inbred lines have been innovating for 100 years[J]. Hereditas, 2016, 38(3): 181-195. DOI: 10.16288/j.yczz.15-430.
[16]	ZHIQIANG P, CUN S, YING J, et al. WZS-pig is a potential donor alternative in corneal xenotransplantation[J]. Xenotransplantation, 2007, 14(6): 603-611. doi: 10.1111/xen.2007.14.issue-6
[17]	LI A, PAN Z, JIE Y, et al. Comparison of immunogenicity and porcine-to-rhesus lamellar corneal xenografts survival between fresh preserved and dehydrated porcine corneas[J]. Xenotransplantation, 2011, 18(1): 46-55. DOI: 10.1111/j.1399-3089.2011.00626.x.
[18]	LIU Y, ZHANG Y, LIANG Q, et al. Porcine endothelial grafts could survive for a long term without using systemic immunosuppressors: an investigation of feasibility and efficacy of xeno-Descemet's stripping automated endothelial keratoplasty from WZS-pig to rhesus monkey[J]. Xenotransplantation, 2018: e12433. DOI: 10.1111/xen.12433.
[19]	PATIENCE C, TAKEUCHI Y, WEISS RA. Infection of human cells by an endogenous retrovirus of pigs[J]. Nat Med, 1997, 3(3): 282-286. doi: 10.1038/nm0397-282
[20]	冯书堂, 马玉媛, 夏颖.中国小型猪内源性反转录病毒研究进展与对策[J].中国畜牧兽医, 2017, 44(4): 1244-1248. DOI: 10.16431/j.cnki.1671-7236.2017.04.043. FENG ST, MA YY, XIA Y. Research progress and countermeasures of porcine endogenous retrovirus in Chinese miniature[J]. Chin Anim Husb Vet Med, 2017, 44(4): 1244-1248. DOI: 10.16431/j.cnki.1671-7236.2017.04.043.
[21]	WU J, MA Y, LV M, et al. Large-scale survey of porcine endogenous retrovirus in Chinese miniature pigs[J]. Comp Immunol Microbiol Infect Dis, 2008, 31(4): 367-371. doi: 10.1016/j.cimid.2007.06.004
[22]	ALBL B, HAESNER S, BRAUN-REICHHART C, et al. Tissue sampling guides for porcine biomedical models[J]. Toxicol Pathol, 2016, 44(3): 414-420. DOI: 10.1177/0192623316631023.
[23]	冯书堂, 李奎, 刘岚, 等.小型猪近交系新品种的培育与开发利用[J].农业生物技术学报, 2015, 23(2): 274-280. DOI: 10.3969/j.issn.1674-7968.2015.02.016. FENG ST, LI K, LIU L, et al. Cultivation and application of miniature pig (sus scrofa) inbred[J]. J Agricult Biotechnol, 2015, 23(2): 274-280. DOI: 10.3969/j.issn.1674-7968.2015.02.016.
[24]	ZHAO Y, XIANG L, LIU Y, et al. Atherosclerosis induced by a high-cholesterol and high-fat diet in the inbred strain of the Wuzhishan miniature pig[J]. Anim Biotechnol, 2018, 29(2): 110-118. DOI: 10.1080/10495398.2017.1322974.
[25]	冯书堂.中国实验用小型猪[M].北京:中国农业出版社, 2011.