PMID- 12088830
OWN - NLM
STAT- MEDLINE
DCOM- 20020911
LR  - 20200409
IS  - 0166-0934 (Print)
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 104
IP  - 2
DP  - 2002 Jul
TI  - Continuous porcine cell lines developed from alveolar macrophages: partial 
      characterization and virus susceptibility.
PG  - 203-16
AB  - 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.
FAU - Weingartl, H M
AU  - Weingartl HM
AD  - NCFAD, CFIA, 1015 Arlington St., Winnipeg, MB, Canada R3E 3M4. 
      hweingartl@inspection.gc.ca
FAU - Sabara, M
AU  - Sabara M
FAU - Pasick, J
AU  - Pasick J
FAU - van Moorlehem, E
AU  - van Moorlehem E
FAU - Babiuk, L
AU  - Babiuk L
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (Antigens, Polyomavirus Transforming)
RN  - I16QD7X297 (Neomycin)
SB  - IM
MH  - Animals
MH  - Antigens, Polyomavirus Transforming/*analysis
MH  - Cell Culture Techniques/methods
MH  - Cell Line
MH  - Cell Survival
MH  - Flow Cytometry
MH  - Freezing
MH  - Macrophages, Alveolar/*cytology/*virology
MH  - Microbial Sensitivity Tests/*methods
MH  - Neomycin/pharmacology
MH  - Simian virus 40/drug effects/growth & development/*physiology
MH  - Swine
MH  - Virus Replication
PMC - PMC7119708
EDAT- 2002/06/29 10:00
MHDA- 2002/09/12 10:01
PMCR- 2002/06/13
CRDT- 2002/06/29 10:00
PHST- 2002/06/29 10:00 [pubmed]
PHST- 2002/09/12 10:01 [medline]
PHST- 2002/06/29 10:00 [entrez]
PHST- 2002/06/13 00:00 [pmc-release]
AID - S016609340200085X [pii]
AID - S0166-0934(02)00085-X [pii]
AID - 10.1016/s0166-0934(02)00085-x [doi]
PST - ppublish
SO  - J Virol Methods. 2002 Jul;104(2):203-16. doi: 10.1016/s0166-0934(02)00085-x.