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 and 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]	WEI Mingbang, BIANBA Qiongda, XIAO Qingqing, DUAN Mengqi, CHAMBA Yangzom, SHANG Peng. Cloning,Bioinformatics and Expression Analysis of CDKN1B Gene in Tibetan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2196-2206.
[2]	ZHANG Fangwei, ZHANG Qi, LEI Liangliang, SUN Wusheng, ZHANG Di, ZHANG Yunpeng, ZHANG Jingbo, WANG Xiuquan, ZHANG Jing, ZHANG Shumin. Polymorphism of HBEGF Gene and Its Association with Reproductive Traits in Songliao Black Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2370-2379.
[3]	LU Yukui, LIU Jiaqi, ZHONG Jiacheng, CHEN Jidang, ZHU Wanjun, ZHANG Yishan, ZHANG Jipei. Isolation,Identification and Pathogenicity Analysis of Avibacterium paragallinarum from Geese [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2562-2572.
[4]	PAN Pengcheng, LEI Zongquan, LI Xian, HU Xiangyun, QIN Qiantao, QIN Zhaoxian, GUAN Zhihui, CHEN Baojian, XIE Bingkun. Bioinformatics Analysis,Eukaryotic Expression Vector Construction and Tissue Expression of MYL2 Gene in Luchuan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1796-1806.
[5]	LIU Hongrun, ZHU Siran, FENG Lingli, ZHANG Kun, YAN Gang, WANG Yubin, ZHANG Shuai, JIANG Shan, XU Di, LAN Ganqiu, LIANG Jing. Cloning,Bioinformatics Analysis and Tissue Expression Localization of CDO1 Gene in Large White Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1352-1363.
[6]	LI Zhijie, ZHAO Ying, MA Xin, WANG Meng, YU Sijiu, WANG Libin, PAN Yangyang. Research Progress on Isolation Methods and Main Reproductive Functions of Exosomes from Animal Follicular Fluid [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1480-1488.
[7]	TIAN Changqing, ZHANG Kunzhong, QI Yumei, SHI Wenjing, DONG Zhijie, ZHANG Haohao, HE Zengwen, ZHI Ji, ZHAO Xuehui, CHONG Qian, XUE Huiwen, GOU Huitian. Isolation,Identification and Biological Characteristics Analysis of a Strain of Listeria monocytogenes from Environment [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1543-1555.
[8]	CAO Yingying, LIANG Liang, ZENG Yirong, ZHONG Qinghua, YUAN Xiaofang, TANG Haibo, LENG Jing. Isolation,Identification and Biological Characteristics of Salmonella from Tupaia belangeri chinensis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1718-1728.
[9]	CHANG Yitong, ZHANG Wei, PENG Yinglin, CHEN Chen. Evolutionary Analysis,Target Gene Prediction and Tissue Expression Analysis of miR-192 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 882-892.
[10]	ZHEN Zhen, WANG Mei, WANG Yimin, HU Debao, ZHANG Linlin, LI Xin, GUO Yiwen, GUO Hong, DING Xiangbin. Effect of RNA Methylation Transfer Enzyme METTL3 on the Proliferation and Myogenic Differentiation of Bovine Skeletal Muscle Satellite Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1025-1036.
[11]	LIU Kexin, WANG Chao, ZHANG Ao, LIU Jiali, TAN Bin, ZHANG Shuqin. Isolation,Identification and Genome Sequence Analysis of Bovine Viral Diarrhea Virus LN-1 Strain [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1150-1159.
[12]	CHEN Dong, ZHANG Yanhao, ZHANG Yang, LI Sen, ZHOU Zutao, LI Zili. Isolation,Identification and Drug Sensitivity of 3 Strains of Riemerella anatipestifer [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1250-1258.
[13]	WANG Bingyi, MA Hongcai, ZOU Minghao, FAN Shijie, YUAN Zhenjie, Labaciren, DONG Hailong, ZENG Jiangyong. Isolation,Identification and Drug Resistance Analysis of Streptococcus from Yak [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 745-753.
[14]	ZHONG Huachen, WANG Lifang, GUO Chenyang, LIU Jialin, SONG Jie. Isolation,Identification and Drug Resistance Analysis of Pathogenic Bacteria from Milk Samples and Environment of Mastitis in Inner Mongolia [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 817-826.
[15]	YIN Yi, LYU Yanqiu, CHEN Xuan, CAO Lipeng, ZHANG Junzheng, JIN Yi. Effects of Capacitation on Sperm Quality and Regulation of Acrosin Inhibitor Levels by Ubiquitin in Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 174-185.