REPETITIVE LOW-VOLUME BLOOD SAMPLING METHOD AS A FEASIBLE MONITORING TOOL IN A MOUSE MODEL OF SEPSIS

Blood-based monitoring of immunoinflammatory and organ function fluctuations is essential in models of critical illness. This is challenging in diseased mice as repetitive blood collection may be harmful and/or affect end points. We studied the influence of daily sampling in acutely septic (days 1-5) mice upon survival and selected hematologic and organ function parameters. In addition, we tested the reliabilty of complete blood cell (CBC) count using resuspended blood cells. Female OF-1 and CD-1 mice underwent cecal ligation and puncture (CLP) and were subdivided into Daily and Day 5 groups. Blood was collected daily for 5 days in the Daily group and only on day 5 post-CLP in the Day 5 group. We tested 20 mu L (both strains) and 35 mu L (OF-1 mice) sampling volumes. The 35-mu L volume simultaneously served to test the CBC reliabilty in resuspended versus unprocessed blood. Daily sampling did not affect the 14-day CLP mortality. Compared with the Day 5 group, daily 35-mu L sampling in OF-1 mice decreased the red blood cell count and hemoglobin concentration by 22% and 23% (P < 0.05). In neither strain did daily 20-mu L sampling affect the red blood cell count, whereas there was a 9% hemoglobin decrease (P < 0.05) in OF-1 mice. Although alanine aminotransferase, lactate dehydrogenase, and glucose levels were comparable, urea significantly increased by 24% in the Daily group (20-mu L volume, OF-1 mice). Interleukin 6, platelets, and white blood cell counts remained unaffected. There was an excellent correlation between regular and resuspended CBC for all cell types (r >= 0.9; slope, >= 0.9), except lymphocytes (r > 0.5; slope, >0.5). This method provides a feasible and safe translation of clinically relevant daily immunomonitoring to the mouse sepsis model.