Cryopreservable neutrophil surrogates. Stored cytoplasts from human polymorphonuclear leukocytes retain chemotactic, phagocytic, and microbicidal function.

Cryopreservation of polymorphonuclear leukocytes (PMN) has largely failed, probably because of their rich content of granular (lysosomal) enzymes. We have been developing granule-poor cytoplasts (anucleate fragments) from PMN which retain motile functions of the parent cell. The two types studied here were induced either by brief heating on surfaces (cytokineplasts) or by discontinuous gradient centrifugation (Ficoll) without heat or drugs (U-cytoplasts). Freshly made, these cytoplasts respond chemotactically to formyl peptide (fMet-Leu-Phe), and they take up and kill roughly half as many Staphylococcus aureus as their (larger, granular) parent PMN. Unlike their parent cells, after cryopreservation both cytoplasts remain chemotactic, and in matched experiments they take up and kill staphylococci with undiminished avidity. These findings are the first indications that PMN cytoplasts suitable for clinical use may be feasible.