Gender dimorphism in differential peripheral blood leukocyte counts in mice using cardiac, tail, foot, and saphenous vein puncture methods

BMC Clinical Pathology

Elahé T Crockett¹  Jessica L Borowicz¹  Diana C Doeing¹ 

[1] Departments of Physiology and Surgery, College of Human Medicine, Michigan State University, East Lansing, MI, USA

关键词: leukocyte counts;    foot vein;    Saphenous;    tail vein;    cardiac puncture;    blood collection;    gender dimorphism;    mice;   

Others  :  1086487
DOI  :  10.1186/1472-6890-3-3

 received in 2003-05-07, accepted in 2003-09-12,  发布年份 2003

【 摘 要 】

Background

In many animal models that investigate the pathology of various diseases, there is a need to monitor leukocyte counts and differentials. However, various researchers use a range of different techniques in male and female laboratory animals to collect such blood variable information. These studies are then compared to one another without consideration of the possibility that different bleeding sites or techniques as well as gender may produce varying results. In light of this, the peripheral blood leukocyte counts and differentials of C57BL/6 male and female mice were determined using four blood-sampling techniques: cardiac, tail, foot, and saphenous vein punctures.

Methods

Blood smears were prepared and stained with Wright-stain for differential cell analysis. The total number of peripheral blood leukocytes was determined with the aid of a hemocytometer. Applying ANOVA and Student t-test analysis made comparisons between groups.

Results

The total leukocyte counts obtained using the cardiac puncture method were significantly lower as compared to the other three blood sources; saphenous, tail and foot. There were no significant differences between leukocyte counts of blood samples collected from the tail, saphenous, and foot. Additionally, no significant differences were observed in total leukocyte counts between male and female mice. Differential analysis showed lymphocytes as the predominant cell type present in the peripheral blood of both male and female mice, comprising 75–90% of the total leukocytes. While no significant differences were observed between male and female differential counts of blood collected from saphenous and tail veins, a significant difference in differential counts of blood obtained via cardiac puncture was observed between the male and female groups, suggesting the role of sex hormones. Further, of the four methods, cardiac puncture appeared to be the fastest and more reliable technique, yielding the maximum blood volume with the least amount of stress being exerted on the sampling site.

Conclusions

This information suggests that in studies concerning leukocyte counts and differentials, the animal gender and the sampling site of blood collection should be kept consistent as to avoid introducing any misleading experimental variation, and that cardiac puncture is the best method of blood collection in mice.

【 授权许可】

   
2003 Doeing et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.

【 图 表 】

Figure 1.

【 参考文献 】

• [1]Angelov O, Schroer RA, Heft S, James VC, Noble J: A comparison of two methods of bleeding rats: the venous plexus of the eye versus the vena sublingualis. J Appl Toxicol 1984, 4:258-60.
• [2]Dameron GW, Weingand KW, Duderstadt JM, Odioso LW, Dierckman TA, Schwecke W, Baran K: Effect of bleeding site on clinical laboratory testing of rats: orbital venous plexus versus posterior vena cava. Lab Anim Sci 1992, 42:299-301.
• [3]Van Herck H, Baumans V, Brandt CJ, Boere HA, Hesp AP, van Lith HA, Schurink M, Beynen AC: Blood sampling from the retro-orbital plexus, the saphenous vein and the tail vein in rats: comparative effects on selected behavioural and blood variables. Lab Anim 2001, 35:131-9.
• [4]Quimby FH, Saxon PA, Goff LG: Total white blood cell counts of peripheral and heart blood samples of the rat. Science 1948, 107:447.
• [5]Quimby FH, Goff LG: Effect of source of blood sample on total white cell count of the rat. Am J Physiol 1952, 170:196-200.
• [6]Sakaki K, Tanaka K, Hirasawa K: Hematological comparison of the mouse blood taken from the eye and the tail. Exper Anim. 1961, 10:14-9.
• [7]Nemzek JA, Bolgos GL, Williams BA, Remick DG: Differences in normal values for murine white blood cell counts and other hematological parameters based on sampling site. Inflamm Res 2001, 50:523-7.
• [8]Smith CN, Neptun DA, Irons RD: Effect of sampling site and collection method on variations in baseline clinical pathology parameters in Fischer-344 rats. II. Clinical hematology. Fundam Appl Toxicol 1986, 7:658-63.
• [9]Jilma B, Eichler HG, Breiteneder H, Wolzt M, Aringer M, Graninger W, Rohrer C, Veitl M, Wagner OF: Effects of 17 beta-estradiol on circulating adhesion molecules. J Clin Endocrinol Metab 1994, 79:1619-24.
• [10]Subandrio AL, Sheldon IM, Noakes DE: Peripheral and intrauterine neutrophil function in the cow: the influence of endogenous and exogenous sex steroid hormones. Theriogenology 2000, 53:1591-608.
• [11]Chernyshov VP, Vodianyk MO, Hrekova SP: Effect of female steroid hormones on expression of adhesion molecules by peripheral blood leukocytes. Fiziol Zh 2002, 48:46-53.
• [12]England JM, Bain BJ: Total and differential leucocyte count. Br J Haematol 1976, 33:1-7.
• [13]Gaunt SD, Pierce KR: Myelopoiesis and marrow adherent cells in estradiol-treated mice. Vet Pathol 1985, 22:403-8.
• [14]Northern AL, Rutter SM, Peterson CM: Cyclic changes in the concentrations of peripheral blood immune cells during the normal menstrual cycle. Proceedings of the Society for Experimental Biology and Medicine. 1994, 207:81-8.
• [15]Sluiter W, Hulsing-Hesselink E, Elzenga-Claasen I, Van Furth R: Method to select mice in the steady state for biological studies. J Immunol Methods 1985, 76:135-43.
• [16]Diehl KH, Hull R, Morton D, Pfister R, Rabemampianina Y, Smith D, Vidal JM, van de Vorstenbosch C: A good practice guide to the administration of substances and removal of blood, including routes and volumes. J Appl Toxicol 2001, 21:15-23.