Document Detail

Large scale isolation of human blood monocytes by continuous flow centrifugation leukapheresis and counterflow centrifugation elutriation for adoptive cellular immunotherapy in cancer patients.
MedLine Citation:
PMID:  8083534     Owner:  NLM     Status:  MEDLINE    
The increasing interest in mononuclear phagocytes for adoptive cellular immunotherapy (ACI) trials in cancer patients led us to define a procedural approach to harvest reproducibly highly purified single-cell suspensions of large numbers of functional human circulating blood monocytes (Mo). A semiclosed counterflow centrifugal elutriation (CCE) system has been developed, using a new large capacity Beckman JE 5.0 rotor with one interchangeable 40 ml or 5 ml separation chamber, to purify Mo from mononuclear cell (MNC) concentrates of healthy donors and cancer patients obtained by continuous flow centrifugation leukapheresis (CFCL). This method does not require a Ficoll density gradient centrifugation step. A total of 115 leukapheresis procedures were carried out in 35 patients and in 30 healthy donors by either Cobe 2997 or Cobe Spectra, with a similar efficiency in MNC apheresis. The average yield per leukapheresis procedure was 5.6 x 10(9) MNC of purity 90-100% (25-45% Mo, 40-65% lymphocytes). The average yields per elutriation procedure (R/O fraction) were 1.1 x 10(9) cells (purity 93% Mo) using the 5 ml separation chamber, and 1.5 x 10(9) cells (purity 91%) using the 40 ml separation chamber, with a respective recovery of 82 +/- 7% and 78 +/- 8% Mo. In vitro analysis of the viability and function of the purified Mo shows that neither morphological integrity nor physiological activity was compromised by this two-step isolation procedure, which additionally provides highly purified human Mo suspensions, in a quantity suitable for ACl of cancer patients.
A Faradji; A Bohbot; M Schmitt-Goguel; J C Siffert; S Dumont; M L Wiesel; Y Piemont; A Eischen; J P Bergerat; J Bartholeyns
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of immunological methods     Volume:  174     ISSN:  0022-1759     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  1994 Sep 
Date Detail:
Created Date:  1994-10-12     Completed Date:  1994-10-12     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  297-309     Citation Subset:  IM    
Département d'Onco-Hématologie, Hôpitaux Universitaires de Strasbourg, France.
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MeSH Terms
Cell Separation / methods
Cell Survival
Centrifugation / methods
Immunity, Cellular
Immunization, Passive
Immunotherapy, Adoptive / methods*
Leukapheresis / methods*
Monocytes / cytology*
Neoplasms / therapy

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