Continuous porcine cell lines developed from alveolar macrophages

生物 病毒学 病毒 猪繁殖与呼吸综合征病毒 非洲猪瘟病毒 水泡性口炎病毒 细胞培养 猪瘟 分子生物学 遗传学
作者
Hana M. Weingartl,Marta Sabara,John Pasick,E van Moorlehem,L. A. Babiuk
出处
期刊:Journal of Virological Methods [Elsevier BV]
卷期号:104 (2): 203-216 被引量:133
标识
DOI:10.1016/s0166-0934(02)00085-x
摘要

Porcine monomyeloid cell lines were established following transfection of primary porcine alveolar macrophage cultures with plasmid pSV3neo, carrying genes for neomycin resistance and SV40 large T antigen. The parental clone 3D4 exhibited a relatively rapid doubling time (25.5 h), high plating efficiency and mixed phenotype with respect to growth on a solid support. Single cell cloning of the 3D4 parent resulted in establishment of several cell lines; three of them designated 3D4/2, 3D4/21 and 3D4/31 were selected for further characterization. All three clones supported the replication of vesicular stomatitis virus (VSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), swine vesicular disease virus (SVDV), swine poxvirus, African swine fever virus (ASFV), herpes simplex virus (HSV), parainfluenza virus, bovine adenovirus (BAV), vaccinia virus (VV), and porcine adenovirus (PAV). Under the conditions tested the cells did not support replication of porcine reproductive and respiratory syndrome virus (PRRSV). The swine myeloid character was confirmed for all three clones by fluorescence activated cell scanning (FACS) analysis using monoclonal antibodies 74-22-15 and DH59B, which recognize the pan-myeloid antigen cluster SWC3a. A subpopulation of each cell line was positive for nonspecific esterase activity and phagocytic activity to varying degrees depending on the media formulation. Cells from all three lines exhibited anchorage dependent growth when maintained in RPMI 1640 medium supplemented with 5–15% fetal bovine serum (FBS) and nonessential amino acids. Propagation in commercially formulated serum free media resulted in colony formation and growth in suspension. The addition of dimethyl sulfoxide (DMSO) or phorbol 12-myristate 13-acetate (PMA) to serum free media restored cell attachment. DMSO was also able to induce expression of CD14 monocyte marker in the 3D4/31 cell line maintained in FBS containing medium, as determined by FACS with monoclonal antibody CAM36A. Supplementation of RPMI medium with 10% porcine serum upregulated the expression of CD14 and induced expression of porcine macrophage markers recognized by antibodies 2B10 and 2G6 (Vet. Immunol. Immunopathol. 74 (2000) 163) in all three cell lines. The porcine myelomonocytic cell lines obtained may have a wide variety of applications in porcine virology and immunology.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
7秒前
Likz完成签到,获得积分10
7秒前
不安的秋白完成签到,获得积分10
9秒前
清新的剑心完成签到 ,获得积分10
10秒前
Yiling完成签到,获得积分10
10秒前
12秒前
氕氘氚完成签到 ,获得积分10
16秒前
Hello应助不安的秋白采纳,获得10
18秒前
糯米团的完成签到 ,获得积分10
19秒前
神勇从波完成签到 ,获得积分10
21秒前
yellow完成签到 ,获得积分10
23秒前
虚幻元风完成签到 ,获得积分10
26秒前
xybjt完成签到 ,获得积分10
29秒前
巴达天使完成签到,获得积分10
35秒前
江三村完成签到 ,获得积分10
35秒前
量子星尘发布了新的文献求助10
53秒前
CyberHamster完成签到,获得积分10
1分钟前
xiaohong完成签到,获得积分10
1分钟前
朱比特完成签到,获得积分10
1分钟前
1分钟前
zmuzhang2019发布了新的文献求助10
1分钟前
onestepcloser完成签到 ,获得积分0
1分钟前
zoe完成签到 ,获得积分10
1分钟前
发嗲的慕蕊完成签到 ,获得积分10
1分钟前
Linson完成签到,获得积分10
1分钟前
顾矜应助赵三岁采纳,获得10
1分钟前
yyy2025完成签到,获得积分10
1分钟前
木雨亦潇潇完成签到,获得积分10
1分钟前
香蕉觅云应助nine2652采纳,获得10
1分钟前
量子星尘发布了新的文献求助10
1分钟前
芳华如梦完成签到 ,获得积分10
1分钟前
1分钟前
1分钟前
1分钟前
土豆丝完成签到 ,获得积分10
1分钟前
琦琦完成签到,获得积分10
2分钟前
zzzz完成签到,获得积分20
2分钟前
GEZIKU完成签到 ,获得积分10
2分钟前
2分钟前
2分钟前
高分求助中
【提示信息,请勿应助】关于scihub 10000
Les Mantodea de Guyane: Insecta, Polyneoptera [The Mantids of French Guiana] 3000
徐淮辽南地区新元古代叠层石及生物地层 3000
The Mother of All Tableaux: Order, Equivalence, and Geometry in the Large-scale Structure of Optimality Theory 3000
Handbook of Industrial Diamonds.Vol2 1100
Global Eyelash Assessment scale (GEA) 1000
Picture Books with Same-sex Parented Families: Unintentional Censorship 550
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 4038039
求助须知:如何正确求助?哪些是违规求助? 3575756
关于积分的说明 11373782
捐赠科研通 3305574
什么是DOI,文献DOI怎么找? 1819239
邀请新用户注册赠送积分活动 892655
科研通“疑难数据库(出版商)”最低求助积分说明 815022