A method for isolation of rat lymphocyte-rich mononuclear cells from lung tissue useful for determination of nucleoside triphosphate diphosphohydrolase activity

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• 作者：Jeandre Augusto dos S. Jaques^a ; ^b ; Jo&#xe3 ; o Felipe Peres Rezer^a ; ^b ; Jader Betsch Ruchel^b ; Jessi&#xe9 ; Gutierres^a ; Andr&#xe9 ; Valle Bairros^a ; Iria Luiza Gomes Farias^a ; Sonia Cristina Almeida da Luz^a ; Claudia de Mello Bertoncheli^b ; Maria Rosa Chitolina Schetinger^a ; Vera Maria Morsch^a ; Daniela Bitencourt Rosa Leal^b ; ^{dbitencourtrosaleal@gmail.com}
• 关键词：Lymphocytes ; Lung ; NTPDase ; Mononuclear cells ; Nucleotides
• 刊名：Analytical Biochemistry
• 出版年：2011
• 出版时间：1 March 2011
• 年：2011
• 卷：410
• 期：1
• 页码：34-39
• 全文大小：681 K

文摘

Methods for the isolation of peripheral blood mononuclear cells (PBMCs) and human lung mononuclear cells (LMCs) have been proposed previously. This study describes a method that allows the separation of lymphocyte-rich LMCs from rats. Trypan blue was applied to determine cell viability. White blood cell and differential cell counts were also performed. Relationships between nucleoside triphosphate diphosphohydrolase (NTPDase, EC 3.6.1.5) activities expressed in milligrams of protein, millions of cells, and millions of viable cells were examined as linear correlations. The lung tissue yielded 82.46 % lymphocytes, 8.6 % macrophages, 2.20 % monocytes, and 1.27 % polymorphonuclear cells (PMNs). In LMCs, a very strong correlation was observed as follows: between NTPDase activity, as determined using ATP or ADP as a substrate, expressed in milligrams of protein and that expressed in millions of cells (r  0.91), between that expressed in milligrams of protein and that expressed in millions of viable cells (r  0.91), and between that expressed in millions of cells and that expressed in millions of viable cells (r  0.98). Based on our results, we affirm that NTPDase activity could be expressed in millions of viable cells, millions of cells, or milligrams of protein.