Biochip assembly and assay method thereof

Abstract

The present invention is directed to a biochip assembly comprising a semi-permeable membrane and an assay method using said biochip assembly for carrying out cell based assays.

Ideally, such a method involves measuring the migration of cells in a channel under the influence of an analyte wherein said cells are separated from said analyte by a semi-permeable membrane and said analyte and/or said cells are subjected to controlled flow conditions.

Description

FIELD OF THE INVENTION[0001]The present invention is directed to a biochip assembly comprising a semi-permeable membrane and a method using said biochip assembly for carrying out cell based assays.BACKGROUND OF THE INVENTION[0002]The ability to monitor migration of biological cells through tight layer of other cells and tissues is crucial for understanding of mechanism of many life threatening diseases and development of modern therapeutic drugs. This migration is typically triggered by the presence of a particular chemical either immobilized on a surface or diffused through a tissue.[0003]In inflammatory conditions, for example, the migration of leukocytes from blood vessels into diseased tissues is crucial to the initiation of normal disease-fighting inflammatory responses. At the same time, this process, known as leukocyte recruitment, is also involved in the onset and progression of debilitating and life-threatening inflammatory and autoimmunne diseases. Thus, the ability to con...

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Benefits of technology

[0056]In another embodiment of this aspect of the invention, the method further comprises coating at least one side of the semi-permeable membrane with one or more substances which effect cell function prior to forming a layer of seeded cells on said semi-permeable membrane. Ideally, such substance promotes adhesion of cells. Such substances may be in any form, such as a gel, liquid etc.

[0057]The method may also comprise a further step of coating the internal bore of the channel prior to use with a subst

Problems solved by technology

However, margination is not commonly accepted as yet in the three step migration process described above.

Although considerable effort has been invested on the study of lymphocyte chemoattractants, they remain poorly characterized relative to monocyte and neutrophil chemoattractants.

However, no molecule identified primarily as a lymphocyte chemoattractive factor has been sequenced and cloned.

This excessive time span is necessary in order to get reasonable statistics of cell migration.No physiological flow can be established in this assay thus is not possible to monitor cells though all stages of leukocyte recruitment.There is no control of the gradient: chemokine diffusion in the body might be different than in vitro as it takes longer to get a cell migration on in vitro assays.Changes in cell morphology during chemotaxis cannot be observed in real-time (because cells transmigrate through the filter).

In addition, Boyden chamber assays cannot readily answer many questions related to the leukocyte migration.

However, in blood even a short persistence of a soluble chemokine gradient is not feasible because the constant flow of plasma removes the soluble chemokines from the site of their production.

However, many of these cells are difficult to grow in vitro and, when grown, are not morphologically similar to in vivo tissue.

However, animal studies are costly, tim

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