Study of body fluid samples using flow cytometry: Six years of experience at the Hospital Universitario San Ignacio - Pontificia Universidad Javeriana Bogota - Colombia

Abstract

Flow cytometry (FCM) was implemented in 2008 at the Pontificia Universidad Javeriana and later at the Hospital Universitario San Ignacio to examine special samples of patients with hematological malignancies and solid tumors other than bone marrow and peripheral blood for diagnosis and monitoring. This study describes the main findings of special sample evaluation over a six-year period. In all, 1070 samples of body fluids from patients with benign and malignant diseases were examined by FCM. These samples were stabilized with TransFix TM and stained with six-color immunophenotyping panels. Samples included cerebrospinal fluid, bronchoalveolar lavage, pleural fluid, pericardial fluid and ascite fluid from patients with acute and chronic leukemia, myelodysplastic syndromes, lymphomas, myeloma, autoimmune diseases, immunodeficiencies and solid tumors, among others. Flow cytometry provided important information for the classification and detection of minimal numbers of tumor cells in leukemia and lymphoma cases. This work represents the first national report describing FCM implementation in special samples for diagnosis and clinical monitoring of patients with malignant and benign pathologies. Consequently, over the last decade, use of commercially available cell stabilization solutions has been recommended. These solutions can preserve PB and cerebrospinal fluid (CSF) samples over time periods greater than one week after collection [4, 8 -9]. This stabilization preserves cellularity and integrity of both cell surface and intracellular antigens [9].

Comparative analysis between cytomorphology and flow cytometry methods in central nervous system infiltration assessment in oncohematological patients

“…The literature is emphatic in recommending the use of these media, as the CSF samples are characteristically hypocellular and may present rapid cell degeneration after collection. 4 , 5 , 6 , 12 However, stabilization media are not available in the laboratory routine at our service. To overcome this issue, we only processed samples collected at our institution, provided they had been received within a maximum period of 4 hours after collection.…”

“…The investigation is usually performed at diagnosis and periodically during the monitoring of the chemotherapeutic treatment of these conditions. 1 , 2 , 3 , 4 , 5 …”

“… 2 , 3 , 6 However, this methodology has certain limitations, such as the scarcity of cells to be evaluated because of the limited volume and the hypocellularity of the CSF samples, in addition to the possible technical difficulty in distinguishing normal from neoplastic cells. 1 , 4 , 5 , 6 , 7 , 8 , 9 , 10 …”