Ferromagnetic Cell and Tissue Culture Microcarriers

Abstract

A porous, collagen coated, ferromagnetic cell culture microcarrier, which is suitable for in vitro cell and tissue culture and which facilitates 3D multicellular construct generation. Also provided is a method for creating batches of microcarriers which have inserted within them magnetite (Fe3O4) in the presence of collagen, thus creating a microcarrier which becomes magnetic in nature when placed in a the presence of a magnetic field and which facilitates cellular adherence (via the collagen coating) for 3D construct development.

Description

CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation of PCT Application No. PCT / US2004 / 023746, filed Jul. 23, 2004, which claims benefit of U.S. Provisional Application No. 60 / 481,126, filed Jul. 23, 2003.BACKGROUND OF THE INVENTION [0002] It has long been established that cells and tissue growing in microgravity behave differently than those on Earth. The ongoing challenge for the experimental study of cell behavior under these conditions has been simulating the environment of microgravity so that complete laboratory studies can be conducted on Earth. This provides the obvious advantages of cost-effectiveness and safety. [0003] To address this issue, NASA developed the bioreactor in the 1980s. Essentially, the bioreactor is a cylindrical vessel equipped with a membrane for gas exchange and ports for media exchange and sampling. As the bioreactor turns, the cells continually fall through the medium yet never hit bottom. Under these conditions, the cells ...

AI Technical Summary

Benefits of technology

[0008] In one embodiment of the present invention a method is provided for developing a three-dimensional cell culture model. This is accomplished by providing a plurality of microcarriers, an upper and lower graphite plate, an adjustable platform (whereby the upper and lower plate are adjustably spaced) and an upper strong magnet. Next at least one cell is adhered to the magnetic microcarriers in a gas-permeable tissue culture bag. The tissue culture bag is then placed between the upper graphite plate and the lower graphite plate, situated below the upper magnet whereby the growing cells are levitated by adjustment of the platform to the appropriate height and are held in suspension via natural magnetic forces stabilized by the graphite plates, thereby allowing the study of three-dimensional cell growth in suspension in a constant and nonrandomized environment. In another embodiment of this method the microcarriers are ferromagnetic microcarriers. The method may also be employed wherein the upper and lower graphite plates are pyrolyric graphite plates. Continued growth of the cellular constructs is made possible by placing the adjustable platform in a standard incubator.

Problems solved by technology

The ongoing challenge for the experimental study of cell behavior under these conditions has been simulating the environment of microgravity so that complete laboratory studies can be conducted on Earth.

Unfortunately however, on Earth the clusters become too large to fall slowly.

In ground based studies, however, it has been difficult to separate these parameters from one another in order to examine the influence of each on three-dimensional cellular growth and function.

Commercially available magnetic beads are either too small for use as microcarriers in cell culture (diameters on the order of <10 um which are not feasible for use as cell culture supports), and / or they lack appropriate surface matrix coating to facilitate cell adherence, a requisite factor to maintain cell-cell interactions for 3D construct development during in vitro culture.

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