Separation, Culture and Identification of Ruminal Smooth Muscle Cells of Xiangdong Black Goats Aged 42 Days

LIU Sile, KANG Jinhe, TAN Zhiliang, WANG Zheng

doi:10.3969/j.issn.1006-267x.2015.10.033

Chinese Journal of Animal Nutrition >

2015 , Vol. 27 >Issue 10: 3249 - 3255

DOI: https://doi.org/10.3969/j.issn.1006-267x.2015.10.033

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Separation, Culture and Identification of Ruminal Smooth Muscle Cells of Xiangdong Black Goats Aged 42 Days

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1. College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China;
2. Key Laboratory of Agro-Ecological Processes in Subtropical Region, Hunan Key Laboratory of Agro-Ecological Processes, Institute of Subtropical Agriculture, The Chinese Academy of Sciences, Changsha 410125, China

Received date: 2015-04-01

Online published: 2015-10-15

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Abstract

The present study was carried out to establish a culture method for ruminal smooth muscle cells of goat.Ruminal smooth muscle tissue of goats (42 days of age) was collected, and ruminal smooth muscle cells was cultured in vitro by the method of collagenase digestion.The morphology of cells at the stages of primary culture and subculture was observed under the optical microscope, the growth curve of the 5th generation and the growth activity curve of generations of ruminal smooth muscle cells were tested using the cell counting method.Meanwhile, the ruminal smooth muscle cells were identified using cell immune fluorescence method, and the expression of α-actin in generations (1st to 11th) cells was detected using the Western blotting.The results showed that the primary cultured ruminal smooth muscle cells of goats obtained by digestion of 0.20% type Ⅱ collagenase began adherent growth after cultured for 1 day, an exponential growth started on the 2nd day resembling a "hill" in appearance, the curve stated plateau phase on the 5th day, and the growth activity of cells reach the highest in the 5th generation.The α-actin in the cytoplasm exhibited a positive reaction by immune fluorescence staining and the protein expressed stable, no significant difference in generations was found (P>0.05).The results demonstrate that ruminal smooth muscle cells of goats were obtained successfully by the methods of 0.20% type Ⅱ collagenase digestion, which provides an ideal cell model for the further study on the effects of nutrients on goats' rumen function.

Key words： goat; rumen; smooth muscle cell; primary culture; identification; α-actin

Cite this article

LIU Sile, KANG Jinhe, TAN Zhiliang, WANG Zheng . Separation, Culture and Identification of Ruminal Smooth Muscle Cells of Xiangdong Black Goats Aged 42 Days[J]. Chinese Journal of Animal Nutrition, 2015 , 27(10) : 3249 -3255 . DOI: 10.3969/j.issn.1006-267x.2015.10.033

References

[1] SO I,KIM K W.Nonselective cation channels activated by the stimulation of muscarinic receptors in mammalian gastric smooth muscle[J].Journal of Smooth Muscle Research,2003,39(6):231-247.

[2] JALILZADEH-AMIN G,MAHAM M,BALIR-NAGHADEH B,et al.Effects of Mentha longifolia essential oil on ruminal and abomasal longitudinal smooth muscle in sheep[J].Journal of Essential Oil Research,2012,24(1):61-69.

[3] 许远靖.β-羟丁酸对皱胃平滑肌细胞氧化应激及收缩相关蛋白表达的影响[D].硕士学位论文.长春:吉林大学,2013.

[4] SWEENEY D,HOLLINS F,GOMEZ E,et al.No evidence for altered intracellular calcium-handling in airway smooth muscle cells from human subjects with asthma[J].BMC Pulmonary Medicine,2015,15:12,doi:10.1186/s12890-015-0009-z.

[5] XU S W,FU J J,CHEN J W,et al.Development of an optimized protocol for primary culture of smooth muscle cells from rat thoracic aortas[J].Cytotechnology,2009,61(1/2):65-72.

[6] 丘钦英,周家国,刘玉洁,等.大鼠脑基底动脉平滑肌细胞培养及功能鉴定[J].中国药理学通报,2007,23(11):1531-1533.

[7] 李影.猪脂肪细胞生物学特性及胰岛素和胆固醇在其生脂过程中的调控作用[D].博士学位论文.杨凌:西北农林科技大学,2006.

[8] 臧元伟,高路,吴栋,等.组织块消化法原代培养大鼠输精管平滑肌细胞及鉴定[J].山东大学学报:医学版,2012,50(8):51-56.

[9] CAIRRÃO E,SANTOS-SILVA A J,ALVAREZ E,et al.Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels[J].In Vitro Cellular & Developmental Biology Animal,2009,45(3/4):175-184.

[10] SUH J H,YOON J S,KIM H W,et al.Adventitial fibroblast abormality in thoracic aortic aneurysms and aortic dissections[J].The Korean Journal of Thoracic and Cardiovascular Surgery,2011,44(6):406-412.

[11] 陈威中,吴满平.人脐动脉内皮细胞——平滑肌细胞共培养模型的建立及鉴定[J].复旦学报:医学版,2009,36(6):768-775.

[12] JANSSEN P H,KIRS M.Structure of the archaeal community of the rumen[J].Applied and Environmental Microbiology,2008,74(12):3619-3625.

[13] DEHORITY B A,TIRABASSO P A.Effect of feeding frequency on bacterial and fungal concentrations,pH,and other parameters in the rumen[J].Journal of Animal Science,2001,79(11):2908-2912.

[14] 辛毅,许秀芳,黄益民,等.乳小鼠心肌成纤维细胞和心肌细胞的分离培养及荧光鉴定[J].新乡医学院学报,2011,28(5):541-547.

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