Immunocytochemical Detection of Breast Cancer Cells: A Comparison of Three Attachment Factors

Abstract

The evaluation of contaminating breast cancer cells in hematopoietic grafts is of considerable importance for monitoring the efficiency of purging procedures. We report a comparison of three systems for the in vitro detection and enumeration of metastatic breast cancer cells. Breast cancer cells from established cell lines were mixed with Daudi cells at dilutions ranging from 1:10 to 1:1,000,000, and a predetermined number were fixed in defined areas on microscope slides coated with one of the following attachment factors: (i) Cell-Tak Cell and Tissue Adhesive, (ii) 0.1% solution of Poly-L-Lysine, or (iii) Cel-Line HTC Super Cured slides. We employed a specificity-proven pancytokeratin antibody (A45-B/B3) and the alkaline phosphatase-antialkaline phosphatase (APAAP) staining technique. In multiple experiments, one breast cancer cell in 1,000,000 Daudi cells could reliably be detected in the Cell-Tak and Cel-Line systems and 1 in 100,000, with the Poly-L-Lysine system. The observed number of seeded cells showed a highly significant correlation with the number of cells seeded (p < 0.0001 in all cases). Finally, we used the Cell-Tak method to evaluate clinical material from various sources: from patients with primary carcinomas of the breast, prechemotherapy, and during various chemotherapeutic regimens, as well as from patients with metastatic disease. The system consistently detected tumor cells in bone marrow samples from these patients. All peripheral blood samples from patients with metastatic disease tested positive at incidences ranging from 5 to 19/10(6) peripheral blood mononuclear cells. This is a simple and reliable technique that allows rapid screening of large cell numbers with high resolution of positive cells.

A neuroinductive biomaterial based on dopamine

“…Further, PCD most likely acted as a dopamine receptor antagonist because the addition of a nontoxic level of free dopamine had no positive effects on PC12 cells. Biomaterials with catechol units are known to affect cell adhesion depending on the structure and composition of the polymer (45,46). To the best of our knowledge, the interaction of catecholcontaining polymers with neurons has not been reported.…”

Immunomagnetic separation for enrichment and sensitive detection of disseminated tumour cells in patients with head and neck SCC

“…A white cell count was obtained for each leucocyte-rich plasma prepared to ensure that 2 10 7 cells were tested for each patient. To assess whether tumour cells were lost into the erythrocyte pellet when leucocyte-rich plasma was prepared, 10 ml of venous blood was spiked with varying numbers of cultured tumour cells; aliquots of the erythrocyte pellet were sedimented onto slides coated with Cell-Tak tissue adhesive (5 g/cm 2 ; Becton Dickinson Inc., Oxford, U.K. 27 ), counterstained, and contaminating tumour cells counted. For some experiments, leucocytes were concentrated by centrifugation at 350 g and analysed without DGS.…”

Automated Capillary-Based Vacuum Pulse-Assisted Instrument for Single-Cell Acquisition and Concurrent Detachment/Adhesion Assay, A-picK

“…5 ). PLL is a well-known coating agent that improves the overall adhesion of cells and was used in numerous studies for efficient culturing of cancer cells, 54,55 fibroblasts, 56 embryonic and induced pluripotent stem cells, 57,58 smooth muscle cells, 59 and many others. 60 Thus, the improved adhesion/attachment to the PLL surface observed in our study was not unexpected.…”