A method for isolation of rat lymphocyte-rich mononuclear cells from lung tissue useful for determination of nucleoside triphosphate diphosphohydrolase activity.
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.
Centro de Ciências Naturais e Exatas, Departamento de Química, Laboratório de Enzimologia Toxicológica, Universidade Federal de Santa Maria, Campus Universitário, Sala 2208, 97105-900, Santa Maria, RS, Brazil.
Jeandre Augusto Dos S
JA
Jaques
João Felipe
JF
Peres Rezer
Jader Betsch
JB
Ruchel
Jessié
J
Gutierres
André Valle
AV
Bairros
Iria Luiza
IL
Gomes Farias
Sonia Cristina
SC
Almeida da Luz
Claudia de
Cd
Mello Bertoncheli
Maria Rosa
MR
Chitolina Schetinger
Vera Maria
VM
Morsch
Daniela Bitencourt Rosa
DB
Leal
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D057075:Enzyme Assays / Q000379:methods*; D007958:Leukocyte Count; D008168:Lung / Q000166:cytology*; D008214:Lymphocytes / Q000166:cytology* / Q000201:enzymology*; D008297:Male; D011755:Pyrophosphatases / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D011755:Pyrophosphatases