Plastic and nonmetallic article shaping or treating: processes – With step of cleaning – polishing – or preconditioning...
Reexamination Certificate
1999-03-12
2001-04-10
Silbaugh, Jan H. (Department: 1732)
Plastic and nonmetallic article shaping or treating: processes
With step of cleaning, polishing, or preconditioning...
C264S219000, C264S240000, C264S299000, C264S337000, C264S138000, C264S233000, C424S093700, C435S178000, C435S182000
Reexamination Certificate
active
06214262
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates generally to producing a three dimensional section of a polymerized liquid. The liquid may contain cells and the polymerization is reversible so that the cells may be re-isolated from the three dimensional section.
2. Description of the Prior Art
Previously liquid monomers have been polymerized into three dimensional slabs by casting agarose between slab gel electrophoresis plates separated by 1-mm thick Teflon spacers. After the gelling of the slab was complete, disks were cored from the slab. Cells such as chondrocytes could be added, for example, in DMEM (Dulbecco's Modified Eagles Medium) mixed with PBS (Dulbecco's phosphate buffered saline) containing low melting temperature agarose. This method is useful for testing the mechanical and electromechanical properties of the disks of chondrocyte/agarose disks and control disks with no cells. However, with the use of this method the polymerization of the slab is not reversible. Thus, the cells can not be re-isolated from the slab after the polymerization to conduct further research and testing on the individual cells, such as mRNA testing, etc.
Alginate, a seaweed derived polymer, allows cells to be maintained in a site that resembles native tissue, thus, it is a preferred media for growing and testing cells. Monomers such as alginate, which are viscous liquids polymerized by ion diffusion. Previously the only known method for polymerizing the alginate was to drop small drops of the alginate from the tip of a syringe into a polymerizing bath containing a polymerizing agent. The cell morphology may remain intact, however, the alginate beads are not suitable for mechanical compression studies.
The objective of the present invention is to provide a reversible diffusion controlled method of polymerizing liquid into solids of various shapes, sizes and thicknesses.
Another objective of the present invention is to maintain cells in culture for tissue engineering purposes.
SUMMARY OF THE INVENTION
In accordance with the present invention, there is provided a method of producing a three dimensional section of polymerized liquid, specifically of a monomer. The method includes the utilization of a molding apparatus. The molding apparatus includes a frame having filter paper on each side thereof and support layers on the outer surfaces of the filter paper. The molding apparatus is clamped together and a liquid monomer is added between the frame and one piece of the filter paper. The filled molding apparatus is placed in a bath of polymerizing agent until the liquid polymerizes to form a three dimensional section. The monomer may include cells.
REFERENCES:
patent: 4172128 (1979-10-01), Thiele
patent: 5334640 (1994-08-01), Desai et al.
patent: 5709854 (1998-01-01), Griffith-Cima et al.
Journal of Orthopaedic Research, vol. 10, No. 6, 1992 “Chondrocytes in Agarose Culture Synthesize a Mechanically Functional Extracellular Matrix”.
Chin Vicki
Grodzinsky Alan
Hung Han Hwa
Ragan Paula
Jones Kenneth M.
Massachusetts Institute of Technology
Samuels Gauthier & Stevens
Silbaugh Jan H.
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