Continuous real time ex vivo epifluorescent video microscopy for the study of metastatic cancer cell interactions with microvascular endothelium
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Recent studies suggest that only endothelium-attached malignant cells are capable of giving rise to hematogenous cancer metastases. Moreover, tumor cell adhesion to microvascular endothelium could be crucial in metastasis predilection to specific organs or tissues. However, the existing in vitro and in vivo techniques do not provide for sufficient delineation of distinct stages of a dynamic multi-step intravascular adhesion process. Here we report the development of an experimental system allowing for prolonged continuous ex vivo real-time observation of malignant cell adhesive interactions with perfused microvessels of a target organ in the context of its original tissue. Specifically, the vasculature of excised dura mater perfused with prostate cancer cells is described. An advantage of this technique is that selected fluorescently labeled tumor cells can be followed along identified vascular trees across the entire tissue specimen. The techniques provide for superior microvessel visualization and allow for uninterrupted monitoring and video recording of subsequent adhesion events such as rolling, docking (initial reversible adhesion), locking (irreversible adhesion), and flattening of metastatic cancer cells within perfused microvasculature on a single cell level. The results of our experiments demonstrate that intravascular adhesion of cancer cells differs dramatically from such of the leukocytes. Within dura microvessels perfused at physiological rate, non-interacting, floating, tumor cells move at velocities averaging 7.2×103 μm/s. Some tumor cells, similarly to leukocytes, exhibit rolling-like motion patterns prior to engaging into more stable adhesive interactions. In contrast, other neoplastic cells became stably adhered without rolling showing a rapid reduction in velocity from 2×103 to 0 μm/s within fractions of a second. The experimental system described herein, while developed originally for studying prostate cancer cell interactions with porcine dura mater microvasculature, offers great flexibility in adhesion experiments design and is easily adapted for use with a variety of other tissues including human.
- Al—Mehdi AB, Tozawa K, Fisher AB et al. Intravascular origin of metastasis from the proliferation of endothelium—attached tumor cells: A new model for metastasis. Nat Med 2000; 6: 100–2. CrossRef
- Orr FW, Wang HH, Lafrenie RM et al. Interactions between cancer cells and endothelium in metastasis. J Pathol 2000; 190: 310–29. CrossRef
- Padera TP, Stoll BR, So PTC et al. Conventional and high—speed intravital multiphoton laser scanning microscopy of microvasculature, lymphatics, and leukocyte—endothelial interactions. Mol Imaging 2002; 1: 9–15. CrossRef
- Cooper CR, Pienta KJ. Cell adhesion and chemotaxis in prostate cancer metastasis to bone: a minireview. Prost Cancer Prost Dis 2000; 3: 6–12. CrossRef
- Glinsky VV, Glinsky GV, Rittenhouse—Olsen K et al. The role of Thomsen—Friedenreich antigen in adhesion of human breast and prostate cancer cells to the endothelium. Cancer Res 2001; 61: 4851–7.
- Glinsky VV, Huflejt ME, Glinsky GV et al. Effects of Thomsen—Friedenreich antigen—specific peptide P–30 on β—galactoside—mediated homotypic aggregation and adhesion to the endothelium of MDAMB–435 human breast carcinoma cells. Cancer Res 2000; 60: 2584–8.
- Lehr JE, Pienta KJ. Preferential adhesion of prostate cancer cells to a human bone marrow endothelial cell line. J Natl Cancer Inst 1998; 90: 118–23. CrossRef
- Scott LJ, Clarke NW, George NJR et al. Interactions of human prostatic epithelial cells with bone marrow endothelium: binding and invasion. Br J Cancer 2001; 84: 1417–23. CrossRef
- Nemeth JA, Harb JF, Barroso U et al. Severe combined immunodeficient—hu model of human prostate cancer metastasis to human bone. Cancer Res 1999; 59: 1987–93.
- Yonou H, Yokose T, Kamijo T et al. Establishment of a novel speciesand tissue—specific model of human prostate cancer in humanized nonobese diabetic/severe combined immunodeficient mice engrafted with human adult lung and bone. Cancer Res 2001; 61: 2177–82.
- Rumbaut RE, Huxley VH. Similar permeability responses to nitric oxide synthase inhibitors of venules from three animal species. Microvascular Res 2002; 64: 21–31. CrossRef
- Rubin MA, Pitzi M, Mucci N et al. Rapid (‘warm’) autopsy study for procurement of metastatic prostate cancer. Clin Cancer Res 2000; 6: 1038–45.
- Lehr H—A, Leunig M, Menger MD et al. Dorsal skinfold chamber technique for intravital microscopy in nude mice. Am J Pathol 1993; 143: 1055–62.
- Rumbaut RE, Harris NR, Sial AJ et al. Leakage responses to L—NAME differ with fluorescent dye used to label albumin. Am J Physiol 1999; 276: H333–9.
- Glinsky GV, Glinsky VV. Apoptosis and metastasis: A superior resistance of metastatic cancer cells to programmed cell death. Cancer Lett 1996; 101: 43–51. CrossRef
- Segre G, Silberberg A. Behavior of microscopic rigid particles in Poiseuille flow. II. Experimental results and interpretation. J Fluid Mech 1962; 14: 136–57. CrossRef
- Lawrence MB, Springer TA. Leukocytes roll on selectin at physiologic flow rates: Distinction from and prerequisite for adhesion through integrins. Cell 1991; 65: 859–73. CrossRef
- Butcher EC. Leukocyte—endothelial cell recognition: Three (or more) steps to specificity and diversity. Cell 1991; 67: 1033–6. CrossRef
- Khaldoyanidi SK, Glinsky VV, Sikora L et al. MDA—MB–435 human breast carcinoma cell homo—and heterotypic adhesion under flow conditions is mediated in part by Thomsen—Friedenreich antigengalectinn–3 interactions. J Biol Chem 2003; 278: 4127–34. CrossRef
- Continuous real time ex vivo epifluorescent video microscopy for the study of metastatic cancer cell interactions with microvascular endothelium
Clinical & Experimental Metastasis
Volume 20, Issue 5 , pp 451-458
- Cover Date
- Print ISSN
- Online ISSN
- Kluwer Academic Publishers
- Additional Links
- dura mater
- prostate carcinoma
- video microscopy
- Industry Sectors
- Author Affiliations
- 1. Department of Physiology, University of Missouri, Columbia, Missouri, USA
- 2. Harry S. Truman Memorial Veterans Hospital, Columbia, Missouri, USA
- 3. USA
- 4. Veterinary Biomedical Sciences, University of Missouri, Columbia, Missouri, USA
- 5. Department ofVeterinary Biomedical Sciences, University of Missouri, Columbia, Missouri, USA
- 6. Department of Biochemistry, University of Missouri, Columbia, Missouri, USA
- 7. Departments of Internal Medicine, USA
- 8. Urology, University of Michigan, Ann Arbor, Michigan, USA