title: Peripheral blood lymphocytes are able to maintain their viability and basic function in normal urine.
abstract: BACKGROUND
  Similar to inflammatory cells, peripheral blood mononuclear cells (PBMCs) can also infiltrate in to kidney and urinary tracts and subsequently excreted by urine. In this study we determined the viability rate and response to phytohemagglutinin-A (PHA) of human PBMCs in normal urine.
  
  
  METHODS
  A number of 1×10(6) ficoll-hypaque isolated PBMCs were dispensed in 1 ml normal urine and 6 molar urea and RPMI-1640+FBS10 % were considered as negative and positive control, respectively. After 20, 60 and 120 minutes the viability of these cells was measured by trypan blue dye exclusion assay. 1×10(5) of PBMCs were isolated from urine and cultured as triplicate in RPMI-1640`supplemented with FBS 10% and PHA for 96hr. MTT assay was performed to determine the PBMCs response to PHA. These experiments were repeated three times independently.
  
  
  RESULTS
  There was no significant difference between the viability rates of the PBMCs incubated in urine and positive control after 20, 60 and 120 minutes. Overall, there was a significant difference in trends of viability rate across the three groups (p<0.05).
  
  
  CONCLUSIONS
  Our results showed that not only PBMCs remained remarkably alive in urine after 120 minutes, but can also respond to PHA up to 60 minutes after incubation in urine. These data open a new avenue in the designation for cell culture-based techniques in urine cell analysis.
authors.affiliation: Department of Immunology Babol University of Medical Sciences, Babol, Iran.
authors.forename: Azin
authors.initials: A
authors.lastname: Aghamajidi
authors.affiliation: Department of Immunology Babol University of Medical Sciences, Babol, Iran.
authors.forename: Hesam
authors.initials: H
authors.lastname: Babaie
authors.affiliation: Department of Immunology Babol University of Medical Sciences, Babol, Iran.
authors.forename: Narges
authors.initials: N
authors.lastname: Amirjamshidi
authors.affiliation: Cellular and Molecular Biology Research Center, Babol University of Medical Science, Babol, Iran.
authors.forename: Zeinab
authors.initials: Z
authors.lastname: Abedian
authors.affiliation: Cellular and Molecular Biology Research Center, Babol University of Medical Science, Babol, Iran.
authors.forename: Hamidreza
authors.initials: H
authors.lastname: Khorasani
authors.affiliation: Cellular and Molecular Biology Research Center, Babol University of Medical Science, Babol, Iran.
authors.forename: Amrollah
authors.initials: A
authors.lastname: Mostafazadeh
keywords: PHA; Urine; lymphocyte activation; lymphocyte lifespan