ASGSB 1998 Annual Meeting Abstracts

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MIGRATION OF LYMPHOCYTES INCUBATED IN CLINOSTATS AND STATIC CULTURES.   A.J. Bergman and K. Zygourakis. Departments of Chemical Engineering and Bioengineering, Rice University, Houston.

The objective of this study was to determine if different mechanical stimuli affect the migration of lymphocytes on surfaces coated with the extracellular matrix protein fibronectin. Jurkat cells (a T lymphocyte cell line) were incubated for 30 days either in a clinostat or in a normal static culture. The cells were then seeded on tissue culture plates that were pre-incubated with solutions containing different concentrations of fibronectin. Remaining binding sites were blocked using bovine serum albumin. Migrating cells were monitored for 8 hours using a digital video microscopy setup and their positions on the plate were determined at 10-minute intervals. From these measurements, the cell trajectories were reconstructed using a nearest neighbor algorithm. Cell locomotion was analyzed using the persistent random walk model for two-dimensional systems to determine the random motility coefficient for the cell population. This parameter is similar to an effective diffusion coefficient and was used to compare the locomotory behavior of different cell populations.

For tissue culture plates coated with fibronectin solutions of 0, 4.3, 21.5, and 43 g/ml, the random motility coefficients of Jurkat cells measured 3-7 hours after cell seeding were 4.1±0.3, 11.3±1.0, 9.6±0.6 and 16.2±2.3 µm²/min respectively. Four hours after cell seeding, Jurkat cells cultured in the clinostat showed a lower random motility coefficient (5.0±0.2 µm²/min) than cells cultured statically (10.0±0.27 µm²/min). Similarly, centrifugation (168g) for 10 minutes prior to seeding decreased cell migration rates for the first 5 hours after cell seeding. These effects, however, were reversible. Seven hours after cell seeding, the random motility coefficients of the clinostated and control cells were not significantly different.

(This work was supported by the NASA grant #NAGW-5007)

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