北京鸭肝间充质干细胞分离、培养及分化潜能研究

doi:10.16431/j.cnki.1671-7236.2019.10.009

Abstract

Abstract: The aim of this study was to study the isolatation,culture,identification and multi-lineage differentiation potential of liver mesenchymal stem cells (LMSCs) in Beijing duck.LMSCs were isolated from 19-day-old healthy Beijing duck embryos by enzymatic digestion and cultured in vitro.The specific markers (CD29,CD44,CD71,CD73,CD90,CD105,CD133 and CD166) of LMSCs were detected by immunofluorescence,RT-PCR and flow cytometry.Because the LMSCs were mixed with some hematopoietic stem cells,CD31 and CD34 which were the specific markers of hematopoietic stem cells were detected.Furthermore,LMSCs were induced to differentiate into adipogenic and osteogenic cells to detect their multi-lineage differentiation potential.The results showed that LMSCs obtained by enzymatic digestion expressed specific markers of mesenchymal stem cells,and the specific markers of hematopoietic stem cells were negative.It was proved that LMSCs were hepatic mesenchymal stem cells,and differentiated into adipocytes and osteoblasts.RT-PCR detection results revealed that adipogenic cells expressed peroxisome proliferator-activated receptor γ (PPAR-γ) and lipoprotein lipase (LPL),osteogenic cells expressed Collogen Ⅰ and Runt-related transcription factor 2 (RUNX2).The strong self-proliferation ability of Beijing duck LMSCs made it possible to make unlimited use of livestock and poultry genetic resources.The strong multi-directional differentiation potential indicated that the level of reprogramming was low,and it had great advantages for the recovery of livestock and poultry germplasm resources.It provided a theoretical basis for tissue regeneration research and opened up a broader platform for the development and utilization of livestock and poultry resources.

Key words: Beijing duck; mesenchymal stem cells; biological characteristics; induced differentiation; liver

CLC Number:

S852.16⁺5

LI Tingting, XU Long, LIANG Jun. Study on Isolation,Culture and Differentiation Potential of Liver Mesenchymal Stem Cells in Beijing Duck[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(10): 2882-2889.

References

[1] WANG Y,YU X,CHEN E,et al.Liver derived human mesenchymal stem cells:A novel therapeutic source liver diseases[J].Stem Cell Research Theraphy,2016,7(1):71.
[2] SCHMELZER E,ZHANG L,BRUCE A,et al.Human hepatic stem cells from fetal and postnatal donors[J].Journal Exprimental Medicine,2007,204(8):1973-1987.
[3] KIDO T,KOUI Y,SUZUKI K,et al.CPM is a useful cell surface marker to isolate expandable bipotential liver progenitor cells derived from human iPS cells[J].Stem Cell Reports,2015,5(4):508-515.
[4] 李阳,张斌斌,彭青,等.体外人原代肝细胞分离与培养及冻存研究[J].现代医院,2017,17(6):867-871. LI Y,ZHANG B B,PENG Q,et al.Study on the isolation,cultivation and cryopreservation of human primary hepatocytes in vitro[J]. Modern Hospital,2017,17(6):867-871.(in Chinese)
[5] 李耀庭,吕建军,周舒雅,等.原代肝细胞分离培养及其作为致癌物预测体外模型应用的进展[J].药物分析杂志,2015,35(7):1134-1139. LI Y T,LYU J J,ZHOU S Y,et al.Research progress on primary hepatocyte isolation,culture and its application as in vitro models to predict potential carcinogens[J]. Chinese Journal of Pharmaceutical Analysis,2015,35(7):1134-1139.(in Chinese)
[6] 尹寿新,周震,马美雪.体外大鼠肝卵圆细胞的长期培养方法[J].中国组织工程研究,2014,18(29):4663-4668. YIN S X,ZHOU Z,MA M X.Long-term culture of hepatic oval cells in rats in vitro[J].Chinese Journal of Tissue Engineering Research,2014,18(29):4663-4668.(in Chinese)
[7] TAKAYAMA K,INAMURA M,KAWABATA K,et al.Generation of metabolically functioning hepatocytes from human pluripotent stem cells by FOXA2 and HNF1alpha transduction[J].Journal of Hepatol,2012,57(3):628-636.
[8] TAKAYAMA K,INAMURA M,KAWABATA K,et al.Efficient generation of functional hepatocytes from human embryonic stem cells and induced pluripotent stem cells by HNF4alpha transduction[J].Molecular Therapy the Journal of the American Society of Gene Therapy,2012,20(1):127-137.
[9] COLTER D C,SEKIYA I,PROCKOP D J.Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells[J]. Proceedings of the National Academy of Sciences,2001,98(14):7841-7845.
[10] 牧仁,边艳超,王凤龙,等.北京油鸡胚胎肝脏间充质干细胞的生物学特性[J].华中农业大学学报,2013,32(6):99-105. MU R,BIAN Y C,WANG F L,et al.Biological characterization of mesenchymal stem cells from Beijing fatty chicken fetal liver[J]. Journal of Huazhong Agricultural University,2013,32(6):99-105.(in Chinese)
[11] TAMAJUSUKU A S,CARRILLO-SEPU[DD(-1]＇[HT5"]LVEDA M A,BRAGANHOL E,et al.Activity and expression of ecto-5'-nucleotidase/CD73 are increased by thyroid hormones in vascular smooth muscle cells[J].Molecular and Cellular Biochemistry,2006,289(1-2):65-72.
[12] FAUSTO N.Liver regeneration and repair:Hepatocytes,progenitor cells and stem cells[J].Hepatology,2004,39(6):1477-1487.
[13] KIM S P,HA J M,YUN S J,et al.Transcriptional activation of perxisome proliferator actiated receptor requires activation of both protein kinase A and Akt during adipocyte differntiation[J].Biochemical & Biophysical Research Communication,2019,399(1):55-59.
[14] ALLAMPALLAM K,CHAKRABORTY J,ROB-INSON J.Effect of as corbic acid and growth factorson collagen metabolism of flexor retinaculum cells from individuals with and without carpal tunnel syndrome[J].Journal of Occupational and Environmental Medicine,2000,42(3):251-259.
[15] GARIMELLA R,BI X,ANDERSON H C,et al.Nature of phosphate substrate as a major determinant of mineral type formed in matrix vesicle mediated in vitro mineralization:An FTIR imaging study[J].Bone,2006,38(6):811-817.
[16] 宋明宇,王蓉,杨勇,等.地塞米松对骨髓间充质细胞增殖及成骨、成脂分化的效应[J].骨科,2017,8(4):302-308. SONG M Y,WANG R,YANG Y,et al.The effect of dexamethasone on proliferation,osteogenic or adipogenic differentiation of bone marrow mesenchymal stem cells[J].Orthopaedics,2017,8(4):302-308.(in Chinese)
[17] RAUCH A,SEITZ S,BASCHANT U,et al.Glucocorticoids suppress bone formation by attenuating osteoblast differentiation via the monomeric glucocorticoid receptor[J].Cell Metabolism,2010,11(6):517-531.
[18] 刘伟,刘萌,朱劲松,等.人骨髓间充质干细胞的体外培养,鉴定及成骨分化[J].中国组织工程研究,2012,16(14):2515-2519. LIU W,LIU M,ZHU J S,et al.In vitro culture,identification and osteogenic differentiation of human bone marrow mesenchymal stem cells[J].Chinese Tissue Engineering Study,2012,16(14):2515-2519.(in Chinese)
[19] ARENDS B,SPEE B,SCHOTANUS B A,et al.In vitro differentiation of liver progenitor cells derived from healthy dog livers[J].Stem Cells and Development,2009,18(2):351-358.
[20] HUNCH M,GEHSRT H,BOXTEL R V,et al.Long-term culture of genome-stable bipotent stem cells from adult human liver[J].Cell,2015,160(1-2):299-312.
[21] LIN J S,ZHOU L,SAGAYARAJ A,et al.Hepatic differentiation of human amniotic epithelial cells and in vivo the rapeutic effect on animal model of cirrhosis[J].Journal of Gastroenterol and Hepatol,2015,30(3):1673-1682.

Study on Isolation,Culture and Differentiation Potential of Liver Mesenchymal Stem Cells in Beijing Duck

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	XU Jiahui, XIE Jiaxin, JIANG Chengkun, ZHANG Sen, YAO Hua, ZHANG Hua. Application of Three-dimensional Reconstruction and 3D Printing Technology in Beagle Dog Liver Model Construction [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2130-2138.
[2]	WANG Leping, PAN Yan, TAN Yizhou, LI Lei, HAN Kaiou, CAO Yajie, LI Jun, ZHOU Yuqing, LIAO Yuying, MA Dongxin, GONG Yu, PENG Hao, WANG Xiaoye. Isolation,Identification and Genome Analysis of Escherichia coli Phage vB_EcoP_GN07 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1522-1531.
[3]	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.
[4]	YAO Jiawei, HUANG Yujie, CHEN Zhisheng, WANG Bingyun, ZHANG Hui. Immunomodulatory Effects of Dimethyl Alpha-ketoglutarate Pretreatment on Canine Adipose-derived Mesenchymal Stem Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1642-1652.
[5]	CHENG Longfei, LIAO Weilian, CHEN Hongmei, JIANG Nansong, FU Qiuling, WAN Chunhe, LIU Rongchang, SHI Shaohua, HUANG Yu, FU Guanghua. Screening and Biological Characteristics of Broad-spectrum Riemerella anatipestifer Phage [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1169-1176.
[6]	GAN Lu, ZHENG Huizhen, NUO Mingdalai, LIU Yan, JIN Min, HE Wenwen, WEN Licui, LI Yongchang, GAILIKE·Bayinchahan. Propagation of Trypanosoma evansi Yili Strain and Cloning, Expression and Bioinformatics Analysis of Its PFR Gene [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 469-478.
[7]	LIU Bin, LI Meng, XI Qianyun, SUN Jiajie, LUO Junyi, ZHANG Yongliang, CHEN Ting. Research Progress on Biological Characteristics of Hermetia illucens L. and Its Application in Poultry Production [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 586-597.
[8]	YIN Yangyan, PAN Yan, LI Xiaoning, LI Jun, LIAO Yuying, LI Changting, WEI Tianchao, PENG Hao. Study on Biological Characteristics and Antibacterial Activity in vitro of a Lactobacillus plantarum Strain [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 666-673.
[9]	WU Tao, QI Yunxia, LIU Wei, ZHU Honglong, HUANG Dongwei, DING Haisheng, ZHAO Huiling, CHENG Guanglong, ZHAO Xiaowei. Effect of Colostrum and Milk Feeding on Hepatic Proteins of Calves Based on Proteomics Analysis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 37-45.
[10]	GAO Yefan, SONG Hanan, WANG Yunan, WU Yue, NIU Ruili, ZONG Xianchun, GUAN Weijun. Study on the Isolation, Culture and Differentiation Potential of Limbal Stem Cells from Beijing You Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3372-3381.
[11]	ZHU Xuedan, PAN Xueqing, MA Junyan, ZHOU Lele, ZHANG Shouquan. Research Progress on the Effects of Prostaglandin F_2α and Its Analogues on the Reproductive Performance of Mammal [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3072-3082.
[12]	LU Wengeng, SI Linqing, TIAN Chunyu, YUAN Siqi, XIE Hexin, ZHOU Jiwei, JIN Jidong, HAN Yinghao. Establishment of Oxidative Stress Model of Bovine Endometrial Epithelial Cells and the Effects of Bovine Adipose-derived Mesenchymal Stem Cells on Cell Migration [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2506-2514.
[13]	ZHOU Yuqing, MA Dongxin, WU Jie, WANG Xiaoye. Isolation,Identification and Characterization Analysis of Broad-spectrum Escherichia coli O157∶H7 CVCC4050 Phage vB_EcoM_GXBP08 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2735-2745.
[14]	SHEN Xingling, DING Kangning, WANG Youcong, ZHANG Yian, LIU Yilei, LIU Hanxiao, TANG Luping, HE Yongming. Study on the Mechanism of Polygonum chinense L. Alleviating Liver Injury Induced by Salmonella Typhimurium in Mice [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2768-2777.
[15]	YANG Zemin, LI Shuang, JIN Zhengyu, LIAO Yixiao, YANG Ying, WEN Ming. Isolation,Identification and Biological Characteristics of a Strain of Lactobacillus plantarum from Carp [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2805-2811.