Human granulocyte isolation by continuous flow centrifugation leukapheresis and counterflow centrifugation elutriation (CFCL/CCE)

Autor: L. J. Shields, T. J. Contreras, John F. Jemionek, J. E. French
Rok vydání: 1979
Předmět:
Zdroj: Transfusion. 19:695-703
ISSN: 1537-2995
0041-1132
DOI: 10.1046/j.1537-2995.1979.19680104095.x
Popis: Human granulocytes (PMNL) isolated by continuous flow centriftagatlon leukapheresis (CFCL) were further purified by counterflow centrifugation elutriation (CCE). Studies were conducted to determine the maximal granulocyte capacity of the 4.5-ml elutriation chamber and to determine the efficiency of granulocyte recovery from CFCL concentrates. The maximal capacity of the elutriation chamber was determined to be 1.30 times 109 granulocytes with a 98.8 per cent granulocyte purity and a 51:1 PMNL/RBC ratio. The efficiency of recovery was 82.2 per cent with a 95.1 per cent granulocyte purity and a 68:1 PMNL/RBC ratio. The CCE flow system was designed to allow continuous elutriation runs without need of breakdown time between runs. In four to five hours, approximately 4 to 5 times 109 granulocytes can be isolated with purity in excess of 95 per cent. In vitro analysis of the viability and function of CFCL/CCE-isolated granulocytes indicates that the morphologic integrity and physiologic function were not comprised as a result of the combined isolation procedure relative to granulocytes isolated by CCE from whole blood samples. Granulocytes were held at 4 to 6 C for 16 to 24 hours in granulocyte storage medium (GSM) plus 20 per cent autologous plasma at concentrations of about 6 times 106 PMNL/ml prior to in vitro analysis.
Databáze: OpenAIRE
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