The autologous mixed lymphocyte response: isolation of mononuclear cell populations by counterflow centrifugal elutriation.
We have used counterflow centrifugal elutriation (CCE) to isolate cell populations for an autologous mixed lymphocyte response (AMLR). Enriched T lymphocyte and monocyte populations were obtained rapidly without the need for cells to first form rosettes with sheep RBC (T cells) or adhere to plastic (monocytes). In 16 separate experiments, peripheral blood (PB) was separated by CCE (9) and/or a plastic adherence and nylon wool column method (9). Viability by all methods was greater than 95% CCE yielded greater than 97% lymphocytes in F20 and greater than 90% lymphocytes in F26. Cells obtained after passage through nylon wool yielded greater than 90% lymphocytes. Both CCE (F34) and plastic adherence techniques yielded greater than 75% monocytes. Similar AMLR response was obtained using either cell separation method. We conclude that mononuclear cells isolated by CCE are responsive in the AMLR and yield comparable data to other more tedious techniques.
Piaskowski
V
V
Medical College of Wisconsin, Department of Pediatrics, Midwest Children's Cancer Center at Children's Hospital of Wisconsin 53233.
Nikaein
A
A
Lauer
S
S
Casper
J T
JT
D000328:Adult; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D002498:Centrifugation; D005069:Evaluation Studies as Topic; D006801:Humans; D066298:In Vitro Techniques; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed