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Clinical and Diagnostic Laboratory Immunology, November 2001, p. 1131-1135, Vol. 8, No. 6
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.6.1131-1135.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Calcein-Acetyoxymethyl Cytotoxicity Assay: Standardization of a Method Allowing Additional Analyses on Recovered Effector Cells and Supernatants

Simona Neri,1 Erminia Mariani,1,2,* Alessandra Meneghetti,2 Luca Cattini,2 and Andrea Facchini1,2

Dipartimento di Medicina Interna e Gastroenterologia, University of Bologna,1 and Laboratorio di Immunologia e Genetica, Istituto di Ricerca Codivilla-Putti IOR,2 Bologna, Italy

Received 2 April 2001/Returned for modification 25 June 2001/Accepted 17 August 2001

Cytotoxicity assays provide an in vitro evaluation of the lytic activity of NK and T cells against tumors or transformed cells. However, none of these methods allow the recovery of cells or supernatants after the assay. We standardized a microcytotoxicity test using calcein-acetoxymethyl (calcein-AM) dye that requires very small quantities of cells while maintaining the same sensitivity as the traditional 51Cr assay. The assay is applicable to resting as well as activated human effector cells and uses different targets such as human cell lines that are adherent or growing in suspension and resistant or sensitive. The most important feature of the method is the possibility of recovering cells and supernatants for additional analyses such as phenotyping and evaluation of soluble factors.

* Corresponding author. Mailing address: Laboratorio di Immunologia e Genetica, Istituto di Ricerca Codivilla-Putti I.O.R., Via di Barbiano 1/10, 40136 Bologna, Italy. Phone: 39051-6366803. Fax: 39051-6366807. E-mail: marianie{at}alma.unibo.it.

Clinical and Diagnostic Laboratory Immunology, November 2001, p. 1131-1135, Vol. 8, No. 6
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.6.1131-1135.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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