Middleton M M, Campbell P A
Department of Medicine, National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado 80206.
J Leukoc Biol. 1989 Nov;46(5):461-6. doi: 10.1002/jlb.46.5.461.
When sterile gelatin sponges are implanted under the skin of a mouse and retrieved 6 hr later, approximately 10(6) neutrophils per mouse, in a 98-99% pure population, may be retrieved by simply squeezing and rinsing the sponges. These neutrophils behave similarly to peritoneal exudate neutrophils in chemotaxis and phagocytosis assays, but are bacteriostatic rather than bactericidal. The sponge method yields substantially more neutrophils than could be obtained by exsanguinating the mouse and isolating blood neutrophils, and a 5-fold purer population than is normally obtained by passage of peritoneal exudate cells over Ficoll. In addition, sponge-elicited neutrophils may be ready for use within a half hour after removal from the mouse, without being exposed to osmotic shock or Ficoll.
将无菌明胶海绵植入小鼠皮下,6小时后取出,通过简单挤压和冲洗海绵,每只小鼠可获得约10⁶个中性粒细胞,纯度达98 - 99%。在趋化性和吞噬作用检测中,这些中性粒细胞的行为与腹膜渗出液中的中性粒细胞相似,但具有抑菌而非杀菌作用。与通过放血处死小鼠并分离血液中性粒细胞相比,海绵法可获得更多的中性粒细胞,且纯度比通过腹膜渗出细胞经聚蔗糖密度梯度离心法获得的通常纯度高5倍。此外,从小鼠体内取出后半小时内,海绵诱导的中性粒细胞即可使用,无需经历渗透压休克或经过聚蔗糖密度梯度离心。