Development and testing of a new disposable sterile device for labelling white blood cells

A. Signore*, A. W. J. M. Glaudemans, G. Malviya, E. Lazzeri, N. Prandini, A. L. Viglietti, E. F. J. De Vries, R. A. J. O. Dierckx

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

7 Citations (Scopus)

Abstract

Aim. White blood cell (WBC) labelling requires isolation of cells from patient's blood under sterile conditions using sterile materials, buffers and disposables under good manufacturing practice (GMP) conditions. Till now, this limited the use of white blood cell scintigraphy (WBC-S) only to well equipped laboratories with trained personnel. We invented, developed and tested a disposable, sterile, closed device for blood manipulation, WBC purification and radionuclide labelling without exposing patient's blood and the operator to contamination risks. This device prototype and a final industrialized device (Leukokit (R)) were tested for WBC labelling and compared to standard procedure. Leukokit (R) was also tested in an international multi-centre study for easiness of WBC purification and labelling.

Methods. On the device prototype we tested in parallel, with blood samples from 7 volunteers, the labelling procedure compared to the standard procedure of the International Society of Radio labeled Blood Elements (ISORBE) consensus protocol with respect to cell recovery, labelling efficiency (LE), cell viability (Trypan Blue test) and sterility (haemoculture). On the final Leukokit (R) we tested the biocompatibifity of all components, and again the LE, erythro-sedimentation rate, cell viability, sterility and apyrogenicity. ACD-A, HES and PBS provided by Leukokit (R) were also compared to Heparin, Dextran and autologous plasma, respectively. In 4 samples, we tested the chemotactic activity of purified WBC against 1 mg/ml of lipopolysaccharide (LPS) and chemotaxis of Tc-99m-HMPAO-labelled WBC (925 MBq) was compared to that of unlabelled cells. For the multi-centre study, 70 labellings were performed with the Leukokit by 9 expert operators and 3 beginners from five centers using blood from both patients and volunteers. Finally, Media-Fill tests were performed by 3 operators on two different days (11 procedures) by replacing blood and kit reagents with bacterial culture media (Tryptic Soy Broth) and testing sterility of aliquots of the medium at the end of procedure.

Results. Tests performed with the prototype showed no significant differences with the standard procedure but a faster and safer approach. Tests performed with the final Leukokit (R) confirmed full biocompatibility, sterility and apyrogenicity of all reagents and plastic ware. Average WBC recovery with Leukokit (R) was comparable to that of the ISORBE protocol (117x10(6)+/- 24x10(6) vs. 132x10(6)+/- 29x10(6) cells, P=not significant). No differences in red blood cells and platelet content were observed. LE was 82% +/- 3% for Leukokit (R) and 65 +/- 5% for control (P=0.0003) being PBS vs autologous plasma the main reason of such difference. Cell viability was always >99.9% in both conditions. Chemotactic tests showed no differences between all Leukokit samples and controls. Haemocultures and Media-Fill tests were always sterile. The procedure was well accepted by expert operators and beginners, with a very fast learning curve (confidence after 2 +/- 2 labellings).

Conclusion. The invented device offers high level of protection to operators and patients. The derived Leukokit (R) is safe and easy to use, and gives a high LE of WBC without affecting cell viability and function. Being a registered closed, sterile medical device, it may allow easier and faster WBC labelling that is not limited to only well equipped laboratories. Also simultaneously labelling of multiple patients is possible.

Original languageEnglish
Pages (from-to)400-408
Number of pages9
JournalQuarterly Journal of Nuclear Medicine and Molecular Imaging
Volume56
Issue number4
Publication statusPublished - Aug-2012

Keywords

  • Leukocytes
  • Radionuclide imaging
  • Infection
  • BONE-INFECTIONS
  • DIAGNOSIS

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