en Nephrology Nephrology Original Article Original Article <p dir="ltr" style="text-align: justify;"><span style="font-family: times new roman;"><span style="color: rgb(0, 0, 255);"><strong><em>Background: </em></strong></span>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.</span></p> <p dir="ltr" style="text-align: justify;"><span style="font-family: times new roman;"><span style="color: rgb(0, 0, 255);"><strong><em>Methods:</em></strong></span> A number of 1&times;10⁶ 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&times;10⁵ of PBMCs were isolated from urine and cultured as triplicate in RPMI-1640`supplemented with FBS 10% and&nbsp; PHA for 96hr. MTT assay was performed to determine the PBMCs response to PHA<span dir="RTL">. </span>These experiments were repeated three times independently.</span></p> <p dir="ltr" style="text-align: justify;"><span style="font-family: times new roman;"><span style="color: rgb(0, 0, 255);"><strong><em>Results:</em></strong></span> There was no significant difference between the viability rates of the PBMCs incubated in urine and positive control after 20, 60 and 120 minutes while after 60 minutes they exhibited 75.6% of reactivity to PHA versus positive control. Overall, there was a significant difference in trends of viability rate across the three groups (p<0.05).</span></p> <p dir="ltr" style="text-align: justify;"><span style="font-family: times new roman;"><span style="color: rgb(0, 0, 255);"><strong><em>Conclusion: </em></strong></span>Our results showed that not only PBMCs remained alive in urine after 120 minutes, but can also respond to PHA up to 60 minutes at a remarkable level. These data open a new avenue in the designation for cell culture-based techniques in urine cell analysis.</span></p> urine, lymphocyte, viability 43 47 http://caspjim.com/browse.php?a_code=A-10-361-1&slc_lang=en&sid=1 Azin Aghamajidi azin_aghamajidi88@yahoo.com 10031947532846004722 10031947532846004722 No Student Research Committee, Babol University of Medical Sciences, Babol, Iran Hesam Babaei khameneh hesam_we5@yahoo.com 10031947532846004723 10031947532846004723 No Student Research Committee, Babol University of Medical Sciences, Babol, Iran Narges Amirjamshidi narges,amirjamshidi@yahoo.com 10031947532846004724 10031947532846004724 No Student Research Committee, Babol University of Medical Sciences, Babol, Iran Zeinab Abedian zeinab_abedian@yahoo.com 10031947532846004725 10031947532846004725 No Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran Hamidreza Khorasani hamid.reza.khorasani@gmail.com 10031947532846004726 10031947532846004726 No Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran Amrollah Mostafazadeh amrolah65@yahoo.com 10031947532846004727 10031947532846004727 Yes Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran