猪骨骼肌卫星细胞体外分离培养研究进展

doi:10.16431/j.cnki.1671-7236.2022.08.009

Abstract

Abstract: Porcine skeletal muscle is an important movement organization of animal body,the main meat source of human beings,and a good model for muscle growth and disease research.After birth,pig needs the participation of muscle satellite cells in the growth and development of skeletal muscle,and the damage repair.Isolation and culture of porcine skeletal muscle satellite cells in vitro is the basis for in-depth study on the growth and development of skeletal muscle and the pathogenesis of diseases,and the premise for molecular function verification at the cellular level.With the deepening of the research on muscle development and pathological molecular mechanism,the technology of isolation and culture of satellite cells of porcine skeletal muscle in vitro has developed rapidly.The longissimus dorsi muscle,hind leg muscle and semitendinosus muscle are commonly used to isolate skeletal muscle satellite cells,and the longissimus dorsi muscle of 1-day-old piglets have the best isolation effect.Enzymes commonly used to isolate skeletal muscle satellite cells include pronase,collagenase,trypsin,collagenase,etc.,and the time of digestion of each enzyme and its combination is different.The best filtration method is 200 mesh +400 mesh combined filtration,and high purity cells can be obtained by three times centrifugation.The usual medium is DMEM/F12+10% fetal bovine serum (FBS)+1% penicillin-streptomycin (P/S).Common markers of skeletal muscle satellite cells include pairing box gene 3(PAX3),PAX7,Myo-genic determinant 5(Myf5),Myf4,Myo-differentiation factor (MyoD),myogenin (MyoG) and so on.In this paper,the isolation,culture and identification of muscle satellite cells of porcine skeletal muscle were reviewed,and the best parameters in each step were sorted out,which provided references for establishing a standardized procedure for isolating porcine skeletal muscle satellite cells,in order to provide theoretical and technical support for muscle development and disease research.

Key words: pigs; muscle satellite cells; isolation; in vitro culture; enzyme digestion method

CLC Number:

S828

YU Zonggang, MA Haiming. Research Progress on Isolation and Culture of Porcine Skeletal Muscle Satellite Cells in vitro[J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(8): 2931-2942.

References

[1] GROENEN M A,ARCHIBALD A L,UENISHI H,et al.Analyses of pig genomes provide insight into porcine demography and evolution[J].Nature,2012,491(7424):393-398.
[2] NAKAMURA A,TAKEDA S.Mammalian models of duchenne muscular dystrophy:Pathological characteristics and therapeutic applications[J].Journal of Biomedicine&Biotechnology,2011,2011:184393-184400.
[3] KLYMIUK N,BLUTKE A,GRAF A,et al.Dystrophin-deficient pigs provide new insights into the hierarchy of physiological derangements of dystrophic muscle[J].Human Molecular Genetics,2013,22(21):4368-4382.
[4] YANG J,LIU H,WANG K,et al.Isolation,culture and biological characteristics of multipotent porcine skeletal muscle satellite cells[J].Cell and Tissue Banking,2017,18(4):513-525.
[5] VERDIJK L B,SNIJDERS T,DROST M,et al. Satellite cells in human skeletal muscle;From birth to old age[J].Age (Dordrecht,Netherlands),2014,36(2):545-547.
[6] FARUP J,RAHBEK S K,RIIS S,et al.Influence of exercise contraction mode and protein supplementation on human skeletal muscle satellite cell content and muscle fiber growth[J].Journal of Applied Physiology,2014,117(8):898-909.
[7] ZHAO X,ZHU R,WANG Y,et al.Differentiation proliferative capacity of skeletal muscle satellite cells from Dapulian and Landrace pigs[J].Italian Journal of Animal Science,2020,19(1):574-585.
[8] HAWKE T,GARRY D.Myogenic satellite cells:Physiology to molecular biology[J].Journal of Applied Physiology,2001,91(2):534-551.
[9] MCCARTHY J J,MULA J,MIYAZAKI M,et al. Effective fiber hypertrophy in satellite cell-depleted skeletal muscle[J].Development (Cambridge),2011,138(17):3657-3666.
[10] LEPPER C,PARTRIDGE T A,FAN C M.An absolute requirement for Pax7-positive satellite cells in acute injury-induced skeletal muscle regeneration[J].Development,2011,138(17):3639-3646.
[11] METZGER K,TUCHSCHERER A,PALIN M,et al. Establishment and validation of cell pools using primary muscle cells derived from satellite cells of pig skeletal muscle[J].In Vitro Cellular&Developmental Biology-Animal,2020,56(3):193-199.
[12] MAURO A.Satellite cell of skeletal muscle fibers[J].The Journal of Biophysical and Biochemical Cytology,1961,9(2):493-495.
[13] KATZ B.The terminations of the afferent nervefibre in the muscle spindle of the frog[J].Philosophical Transactions of the Royal Society of London,1961,243(703):221-240.
[14] GIBSON M C,SCHULTZ E.Age-related differences in absolute numbers of skeletal muscle satellite cells[J].Muscle&Nerve,1983,6(8):574-580.
[15] YOSHIOKA K,KITAJIMA Y,OKAZAKI N,et al.A modified pre-plating method for high-yield and high-purity muscle stem cell isolation from human/mouse skeletal muscle tissues[J].Frontiers in Cell and Developmental Biology,2020,8:793.
[16] DOUMIT M E,MERKEL R A.Conditions for isolation and culture of porcine myogenic satellite cells[J].Tissue Cell,1992,24(2):253-262.
[17] BEKOFF A,BETZ W.Properties of isolated adult rat muscle fibres maintained in tissue culture[J].Journal of Physiology,1977,271(2):537-547.
[18] WILSCHUT K J,TJIN E P M,HAAGSMAN H P,et al.Approaches to isolate porcine skeletal muscle stem and progenitor cells[J].Protocol Exchange,2015.DOI:10.5281/zenodO.15596.
[19] BRINKMEIER H,SEEWALD M J,EICHINGER H M,et al.Culture conditions for the production of porcine myotubes and myoballs[J].Journal of Animal Science,1993,71(5):1154-1160.
[20] 刘月光,史新娥,沈清武,等.利用单根肌纤维法分离和培养猪骨骼肌卫星细胞及其成肌诱导分化[J].农业生物技术学报,2011,19(5):856-863. LIU Y G,SHI X E,SHEN Q W,et al.Isolation and culture of porcine skeletal muscle satellite cells by single muscle fiber and their muscle differentiation[J].Journal of Agricultural Biotechnology,2011,19(5):856-863.(in Chinese)
[21] 罗桂芬,文旭辉,杨公社.猪肌卫星细胞的分离培养及鉴定[J].细胞与分子免疫学杂志,2006,6:823-825. LUO G F,WEN X H,YANG G S.Isolation,culture and identification of porcine muscle satellite cells[J].Chinese Journal of Cellular and Molecular Immunology,2006,6:823-825.(in Chinese)
[22] 吕晓,朱海鲸,曹晖,等.猪肌肉卫星细胞的分离培养及生物学特性鉴定[J].中国兽医学报,2011,31(10):1480-1484. LYU X,ZHU H J,CAO H,et al.Isolation,culture and identification of biological characteristics of porcine muscle satellite cells[J].Chinese Journal of Veterinary Science,2011,31(10):1480-1484.(in Chinese)
[23] SEBASTIAN S,GOULDING L,KUCHIPUDI S V,et al.Extended 2D myotube culture recapitulates postnatal fibre type plasticity[J].BMC Cell Biology,2015,16(1):23-32.
[24] MIERSCH C,STANGE K,ROENTGEN M.Effects of trypsinization and of a combined trypsin,collagenase,and DNase digestion on liberation and in vitro function of satellite cells isolated from juvenile porcine muscles[J].In Vitro Cellular&Developmental Biology-Animal,2018,54(6):406-412.
[25] LI B,LI P,HUANG R,et al.Isolation,culture and identification of porcine skeletal muscle satellite cells[J].Asian-Australasian Journal of Animal Sciences,2015,28(8):1171-1177.
[26] MIERSCH C,STANGE K,ROENTGEN M. Separation of functionally divergent muscle precursor cell populations from porcine juvenile muscles by discontinuous percoll density gradient centrifugation[J].BMC Cell Biology,2018,19(1):2-13.
[27] CHOI K,YOON J W,KIM M,et al.Optimization of culture conditions for maintaining pig muscle stem cells in vitro[J].Food Science of Animal Resources,2020,40(4):659-667.
[28] STANGE K,AHRENS H E,VON MALTZAHN J,et al.Isolation and ex vivo cultivation of single myofibers from porcine muscle[J].In Vitro Cellular&Developmental Biology-Animal,2020,56(8):585-592.
[29] LI J,SU T,ZOU C,et al.Long non-coding RNA H19 regulates porcine satellite cell differentiation through miR-140-5p/SOX4 and DBN1[J].Frontiers in Cell and Developmental Biology,2020,8:518724.
[30] PALIN M F,LAPOINTE J,GARIÉPY C,et al. Characterisation of intracellular molecular mechanisms modulated by carnosine in porcine myoblasts under basal and oxidative stress conditions[J].PLoS One,2020,15(9):e239496.
[31] VAUGHN M A,PHELPS K J,GONZALEZ J M. In vitro supplementation with the porcine plasma product,betaGRO^®,stimulates activity of porcine fetal myoblasts and neonatal satellite cells in a divergent manner[J].Animal,2018,12(9):1912-1920.
[32] KHATRI O S,VAUGHN M A,PHELPS K J,et al.In vitro betaGRO^® supplementation stimulates myogenesis of porcine fetal myoblasts and porcine satellite cells in a divergent manner[J].Journal of Animal Science,2017,954:157.
[33] YAN J,GAN L,YANG H,et al.The proliferation and differentiation characteristics of co-cultured porcine preadipocytes and muscle satellite cells in vitro[J].Molecular Biology Reports,2013,40(4):3197-3202.
[34] DING S,WANG F,LIU Y,et al.Characterization and isolation of highly purified porcine satellite cells[J].Cell Death Discovery,2017,3(2017):17003-17013.
[35] LEWIS F C,HENNING B J,MARAZZI G,et al.Porcine skeletal muscle-derived multipotent PW1(pos)/Pax7(neg) interstitial cells:Isolation,characterization,and long-term culture[J].Stem Cells Translational Medicine,2014,3(6):702-712.
[36] KIM K,KIM D,MIN Y,et al.Myogenic regulatory factors are key players in determining muscle mass and meat quality in Jeju native and Berkshire pigs[J].Veterinary Medicine and Science,2021,7(3):735-745.
[37] WANG X,YANG W,YANG Z,et al.The differential proliferative ability of satellite cells in Lantang and Landrace pigs[J].PLoS One,2012,7(3):e32537.
[38] DOUMIT M E,MERKEL R A.Conditions for isolation and culture of porcine myogenic satellite cells[J].Tissue&Cell,1992,24(2):253-262.
[39] BAQUERO-PEREZ B,KUCHIPUDI S V,NELLI R K,et al.A simplified but robust method for the isolation of avian and mammalian muscle satellite cells[J].BMC Cell Biology,2012,13(1):16-27.
[40] CHEN Y,ZHU H,MCCAULEY S R,et al. Diminished satellite cell fusion and S6K1 expression in myotubes derived from skeletal muscle of low birth weight neonatal pigs[J].Physiological Reports,2017,5(3):e13075.
[41] DENG B,ZHANG F,WEN J,et al.The transcriptomes from two adipocyte progenitor cell types provide insight into the differential functions of MSTN[J].Genomics,2020,112:3826-3836.
[42] KHATRI O S,VAUGHN M A,PHELPS K J,et al. In vitro betaGRO^® supplementation stimulates myogenesis of porcine fetal myoblasts and porcine satellite cells in a divergent manner[J].Journal of Animal Science,2017,954:157.
[43] MIERSCH C,STANGE K,HERING S,et al. Molecular and functional heterogeneity of early postnatal porcine satellite cell populations is associated with bioenergetic profile[J].Scientific Reports,2017,7(1):45052.
[44] STANGE K,MIERSCH C,SPONDER G,et al.Low birth weight influences the postnatal abundance and characteristics of satellite cell subpopulations in pigs[J].Scientific Reports,2020,10(61491):6149-6162.
[45] SUN Y,QIN J,LIU S,et al.PDGFR alpha regulated by miR-34a and FoxO1 promotes adipogenesis in porcine intramuscular preadipocytes through Erk signaling pathway[J].International Journal of Molecular Sciences,2017,18(11):2424-2436.
[46] CHU W,WEI W,HAN H,et al.Muscle-specific downregulation of GR levels inhibits adipogenesis in porcine intramuscular adipocyte tissue[J].Scientific Reports,2017,7(510):510-522.
[47] MILNER D J,BIONAZ M,MONACO E,et al. Myogenic potential of mesenchymal stem cells isolated from porcine adipose tissue[J].Cell and Tissue Research,2018,372(3):507-522.
[48] WANG S,SUN Y,REN R,et al.H3K27me3 depletion during differentiation promotes myogenic transcription in porcine satellite cells[J].Genes,2019,10(2313):231-243.
[49] REDSHAW Z,LOUGHNA P T.Adipogenic differentiation of muscle derived cells is repressed by inhibition of GSK-3 activity[J].Frontiers in Veterinary Science,2018,5:110-115.
[50] LV W,JIN J,XU Z,et al.lncMGPF is a novel positive regulator of muscle growth and regeneration[J].Journal of Cachexia Sarcopenia and Muscle,2020,11(6):1723-1746.
[51] QIU K,XU D,WANG L,et al.Association analysis of single-cell RNA sequencing and proteomics reveals avital role of Ca²⁺ signaling in the determination of skeletal muscle development potential[J].Cells,2020,9(4):1045-1065.
[52] WANG S,XU X,LIU Y,et al.RIP-Seq of EZH2 identifies TCONS-00036665 as a regulator of myogenesis in pigs[J].Frontiers in Cell and Developmental Biology,2021,8:618617.
[53] WANG D,GAO C Q,CHEN R Q,et al.Focal adhesion kinase and paxillin promote migration and adhesion to fibronectin by swine skeletal muscle satellite cells[J].Oncotarget,2016,7(21):30845-30854.
[54] RICCOBONO D,AGAY D,FRANCOIS S,et al. Contribution of intramuscular autologous adipose tissue-derived stem cell injections to treat cutaneous radiation syndrome:Preliminary results[J].Health Physics,2016,111(2):117-126.
[55] YU Q P,FENG D Y,HE X J,et al.Effects of a traditional Chinese medicine formula and its extraction on muscle fiber characteristics in finishing pigs,porcine cell proliferation and isoforms of myosin heavy chain gene expression in myocytes[J].Asian-Australasian Journal of Animal Sciences,2017,30(11):1620-1632.
[56] CHEN X,LUO Y,HUANG Z,et al.Akirin2 regulates proliferation and differentiation of porcine skeletal muscle satellite cells via ERK1/2 and NFATc1 signaling pathways[J].Scientific Reports,2017,7:45156.
[57] HOU L,XU J,JIAO Y,et al.miR-27b promotes muscle development by inhibiting MDFI expression[J].Cellular Physiology and Biochemistry,2018,46(6):2271-2283.
[58] LI L,CHENG X,CHEN L,et al.Long noncoding ribonucleic acid MSTRG.59589 promotes porcine skeletal muscle satellite cells differentiation by enhancing the function of PALLD[J].Frontiers in Genetics,2019,10:1220.
[59] CHEN X,XIANG L,JIA G,et al.Leucine regulates slow-twitch muscle fibers expression and mitochondrial function by Sirt1/AMPK signaling in porcine skeletal muscle satellite cells[J].Animal Science Journal,2019,90(2):255-263.
[60] JIN C,ZHANG Z,SONG Z,et al.mTORC1-mediated satellite cell differentiation is required for lysine-induced skeletal muscle growth[J].Journal of Agricultural and Food Chemistry,2020,68(17):4884-4892.
[61] HONG Q D,XU G,HOU L,et al.microRNA-22 inhibits proliferation and promotes differentiation of satellite cells in porcine skeletal muscle[J].Journal of Integrative Agriculture,2020,19(1):225-233.
[62] SONG Z,JIN C,YE M,et al.JAK2-STAT3 pathway mediated satellite cell apoptosis to govern skeletal muscle growth with lysine[J].Journal of Animal Science,2020,984:334.
[63] YANG R,LIU Y,CHENG Y,et al.Effects and molecular mechanism of single-nucleotide polymorphisms of MEG3 on porcine skeletal muscle development[J].Frontiers in Genetics,2021,12:607910-607921.
[64] DOU M,YAO Y,MA L,et al.The long noncoding RNA MyHC IIA/X-AS contributes to skeletal muscle myogenesis and maintains the fast fiber phenotype[J].Journal of Biological Chemistry,2020,295(15):4937-4949.
[65] CHEN X,ZHAO C,DOU M,et al.Deciphering the miRNA transcriptome of Rongchang pig longissimus dorsi at weaning and slaughter time points[J].Journal of Animal Physiology and Animal Nutrition,2020,104(3):954-964.
[66] WANG X Y,CHEN X L,HUANG Z Q,et al. microRNA-499-5p regulates porcine myofiber specification by controlling Sox6 expression[J].Animal,2017,11(12):2268-2274.
[67] SUN W,HE T,QIN C,et al.A potential regulatory network underlying distinct fate commitment of myogenic and adipogenic cells in skeletal muscle[J].Scientific Reports,2017,7:44133-44146.
[68] ZHANG X,SUN W,HE L,et al.Global DNA methylation pattern involved in the modulation of differentiation potential of adipogenic and myogenic precursors in skeletal muscle of pigs[J].Stem Cell Research&Therapy,2020,11(1):536-550.
[69] BURTON N M,VIERCK J,KRABBENHOFT L,et al. Methods for animal satellite cell culture under a variety of conditions[J].Methods in Cell Science,2000,22(1):51-61.
[70] TAN Y,JIN Y,WANG S,et al.The RNA surveillance factor UPF1 regulates the migration and adhesion of porcine skeletal muscle satellite cells[J].Journal of Muscle Research and Cell Motility,2020,42(2):203-217.
[71] LEWIS F C,HENNING B J,MARAZZI G,et al. Porcine skeletal muscle-derived multipotent PW1pos/Pax7neg interstitial cells:Isolation,characterization,and long-term culture[J].Stem Cells Translational Medicine,2014,3(6):702-712.
[72] JIAO Y,HUANG B,CHEN Y,et al.Integrated analyses reveal overexpressed Notch1 promoting porcine satellite cells'proliferation through regulating the cell cycle[J].International Journal of Molecular Sciences,2018,19(1):271-300.
[73] WANG S,XU X,LIU Y,et al.RIP-Seq of EZH2 identifies TCONS-00036665 as a regulator of myogenesis in pigs[J].Frontiers in Cell and Developmental Biology,2020,8:618617.
[74] 孙红梅,杨公社,孙超.猪前体脂肪细胞与肌卫星细胞体外联合培养[J].农业生物技术学报,2007,15(4):617-621. SUN H M,YANG G S,SUN C.Co-culture of preadipocytes and myogenic satellite cells in porcine[J].Journal of Agricultural Biotechnology,2007,15(4):617-621.(in Chinese)
[75] LI Y,LI F,LIN B,et al.Myokine IL-15 regulates the crosstalk of co-cultured porcine skeletal muscle satellite cells and preadipocytes[J].Molecular Biology Reports,2014,41(11):7543-7553.
[76] GASTER M,BECK-NIELSEN H,SCHRODER H D.Proliferation conditions for human satellite cells.The fractional content of satellite cells[J].APMIS:Acta Pathologica,Microbiologica,et Immunologica Scandinavica,2001,109(11):726-734.
[77] WILSCHUT K J,HAAGSMAN H P,ROELEN B A.Extracellular matrix components direct porcine muscle stem cell behavior[J].Experimental Cell Research,2010,316(3):341-352.
[78] DUMONT N A,BENTZINGER C F,SINCENNES M,et al. Satellite cells and skeletal muscle regeneration[J].Comprehensive Physiology,2015,5(3):1027-1059.
[79] FUKADA S,UEZUMI A,IKEMOTO M,et al. Molecular signature of quiescent satellite cells in adult skeletal muscle[J].Stem Cells,2007,25(10):2448-2459.
[80] GNOCCHI V F,WHITE R B,ONO Y,et al.Further characterisation of the molecular signature of quiescent and activated mouse muscle satellite cells[J].PLoS One,2009,4(4):e5205.
[81] GAO C Q,XU Y L,JIN C L,et al.Differentiation capacities of skeletal muscle satellite cells in Lantang and Landrace piglets[J].Oncotarget,2017,8(26):43192-43200.
[82] MURRAY R L,ZHANG W,IWANIUK M,et al.Dietary tributyrin,an HDAC inhibitor,promotes muscle growth through enhanced terminal differentiation of satellite cells[J].Physiological Reports,2018,6(10):e13706.
[83] LI L,CHEN Y,NIE L,et al.MyoD-induced circular RNA CDR1as promotes myogenic differentiation of skeletal muscle satellite cells[J].Biochimica et Biophysica Acta-Gene Regulatory Mechanisms,2019,1862(8):807-821.
[84] VAUGHN M A,PHELPS K J,GONZALEZ J M. In vitro supplementation with the porcine plasma product,betaGRO^®,stimulates activity of porcine fetal myoblasts and neonatal satellite cells in a divergent manner[J].Animal,2018,12(9):1912-1920.
[85] WANG S,SUN Y,REN R,et al.H3K27me3 depletion during differentiation promotes myogenic transcription in porcine satellite cells[J].Genes,2019,10(3):231-243.

Research Progress on Isolation and Culture of Porcine Skeletal Muscle Satellite Cells in vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	DAI Lin, HUAI Zhifang, WANG Baowei, ZHANG Ming'ai, FAN Wenlei, ZHANG Jing, WANG Binghan, QI Guofeng. Isolation, Identification and Biological Properties and Safety of A Lactiplantibacillus plantarum Strain [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3070-3083.
[2]	LU Chong, WANG Yuyan, FU Han, LI Tongyang, MIAO Ronghao, LU Yabin, LI Jianlong, LIU Jianhua, GUO Qingyong, MAI Zhanhai, KUANG Ling. Isolation and Identification of the Main Pathogenic Bacteria of Aborted Yili Foals and Their Pathogenicity [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3344-3358.
[3]	YANG Quan, LI Xiao, YAN Zunqiang, WANG Pengfei, HUANG Xiaoyu, GAO Xiaoli, YANG Qiaoli, GUN Shuangbao, YANG Jiaojiao. Cloning,Bioinformatics Analysis and Tissue Expression of CXCL12 Gene in Hezuo Pigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2482-2493.
[4]	REN Hao, ZHU Yixuan, CHAO Tingting, WANG Xiaoyi, LU Shaoxiong, YANG Yongli, CHEN Qiang, LI Mingli. Identification and Functional Prediction of lncRNA in Longissimus Dorsi Muscle of Saba Pigs with Different Growth Rates [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2494-2505.
[5]	WANG Zhongfa, LIU Yanchen, YAN Yan, LI Minjuan, HE Yunan, GUAN Weijun, LIU Wenzhong. Study on Isolation Culture and Biological Characteristics of Pancreatic Mesenchymal Stem Cells in Simmental Cattle [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2519-2530.
[6]	CAO Lihua, LI Huali, REN Huibo, LUO Baoming, LIU Yingying, CUI Qingming, DENG Yuan, ZHU Ji, HU Xionggui, LUO Jianhui, ZUO Jianbo, CHEN Chen, PENG Yinglin. Effects of Gender and Slaughter Weight on Carcass and Meat Quality of Guangyi Black Pigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2612-2625.
[7]	LIU Jiayi, WU Hua, SHEN Tong, WANG Kailong, WANG Wensheng, CHEN Zixin. Effects of Extract of Lycium ruthenicum Murr on Growth Performance,Slaughter Performance,Antioxidant Function and Meat Quality of Bamei Ternary Pigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2637-2649.
[8]	ZHAO Yilong, TANG Na, JING Changhua, XU Qingqing, YIN Xiusheng, WANG Haiming, SUN Jing, LIN Panpan, DONG Lin, LIU Jishan, CAO Rongfeng. Isolation,Identification and Biological Characteristics of a Strain of Lumpy Skin Disease Virus [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2790-2799.
[9]	YANG Qinhong, YANG Jia, DUAN Wenjie, ZHANG Yongxian, ZHU Youshuai, YANG Shaohong, HE Cheng, YIN Hongbin, LI Suhua. Isolation and Identification of a High-temperature Tolerant Bacillus thermoamylovorans and Optimization of Its Enzyme Production Conditions [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2839-2850.
[10]	QIN Xiyu, LIU Xiaoxue, CHAI Yi, LAI Mengxuan, REN Xiaomin, ZHANG Depeng, ZHANG Peng, LIJuntao, LI Yixuan, WANG Ran, HAO Yanling, WU Huijuan, WANG Xiaoyu. Establishment and Evaluation of Double Cannulas Model of Duodenal and Terminal Ileum of Bama Minipigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2078-2087.
[11]	LI Jian, WEI Yanan, LI Falei, ZHANG Huilin, LI Dongwei, LIU Yong, XU Gaoxiao. Research Progress on the Regulation of lncRNA on the Development of Skeletal Muscle Satellite Cells [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2187-2197.
[12]	JIA Yuxuan, LI Yaojiang, ZENG Guanghu, SHEN Xiangyu, GONG Ting. Isolation,Culture and Identification of Testicular Spermatogenic Cells from Congjiang Xiang Pig [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2198-2207.
[13]	LI Tianxiu, LI Xinpeng, DONG Xinxing, LAN Guoxiang, YAN Dawei, ZHU Jiawei. Amplification,Sequence Analysis and Tissue Expression Study of HMOX2 Gene in Lijiang Pigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2219-2231.
[14]	WU Peng, YUAN Yang, YANG Yunyun, FENG Yi, WANG Yan, YANG Ying, CHEN Jiangfeng, JIANG Haibo, WEN Ming. Diagnosis and Pathogen Identification of a Case of Mixed Infection of Duck Plague Virus and Pasteurella multocida in Breeding Goose [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2253-2265.
[15]	WU Yue, SHI Xingya, ZHANG Shuai, ZHAO Yunhuan, GUO Liming, ZUO Yuzhu, FAN Jinghui. Isolation and Identification of Porcine Pseudorabies Virus and Genetic Evolution Analysis of gE and gC Genes [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2266-2277.