Trans-membrane co-culture insert and method for using

Abstract

An insert for trans-membrane co-culture includes a hollow housing with a passageway therethrough, a first end, a second end and a center section intermediate the first and second end. The passageway has an inside diameter at the first end and the second end that is larger than the inside diameter of the passageway at the center section so that the passageway tapers from each end to the center section. The center section includes a porous membrane having a first surface and a second surface attached across and obstructing the passageway. The membrane forms a first receptacle from the passageway toward the first end and forms a second receptacle from the passageway toward the second end. A method for using the insert of the present invention to grow cells on both sides of a porous membrane includes horizontally submerging the insert in liquid medium so that the air in the receptacles is displaced by the medium. The insert is then tipped to an upright position while maintaining one end below the surface of the liquid. The insert is stood upright on the bottom of the well and a first cell suspension introduced in to the first receptacle. After allowing the first cell suspension to form a layer of cells on the first surface of the membrane the insert is removed, drained and again horizontally immersed in liquid medium so that the receptacles are filled. The insert is then returned to vertical while maintaining the first end below the surface of the medium, stood on the first end and a second cell suspension is introduced into the second receptacle. After suitable incubation, the second cell population forms a layer on the second surface of the membrane.