US 5,134,070 A · Method and device for cell cultivation on electrodes

Abstract

A method and device are disclosed for culturing of adherent cell monolayer cultures on an electrode surface and subjecting the cells to an electrical field. The device may generally comprises cell culture dish, for example a petri-type dish, having a bottom, with an electrically conductive, optically transparent coating amenable to cell adhesion on the upper surface of the bottom. A metal electrode may contact the underside of the coating and is connected to a source of electrical power. The method involves use of the device for culturing the cells under the intermittent or continuous influence of an electric field, or during the establishment of an electrical field or potential. Further, a method and device are disclosed for the sensing of cell poration, and the use of this information to automate the process of electroporation. Further yet, a device and method are disclosed for use in assisting in the time-correlated mapping of the sequence of serial and parallel gene replication.

Claims22

Dependent claims

Apparatus for the transfection of exogenous material into in situ adherent cells by means of electroporation while said cells are adherent to an electrode surface, said apparatus comprising:

a chamber means wherein in situ adherent cells may be subjected to an electrical field or sufficient intensity to cause poration of cells while the cells are adherent to an electrode surface;
said chamber means characterized by having at least one internal surface being an electrically conductive surface conductive to cell adhesion and growth forming a first electrode having a first surface a second internal face formed of an electrical conductor forming a second or counter electrode;
and means for supporting said counter electrode in close proximity to said surface having cells adherent thereon;
fluid retaining means for maintaining cells submerged in an appropriate medium while adherent to said surface;
means for applying an electric potential or electrical ionizing source to said first surface and said counter electrode;
means for detecting the poration of cells and converting said detected poration into a modulating signal;
means for retaining a fluid in contact with cells, said first surface, said counter-electrode, and said sensing means;
and
means for controlling by said modulating signal, said electrical potential or electrical ionizing source.

6 more elements

Method of transfecting exogenous material into in situ adherent cells by means of electroporation while said cells are adherent to an electrode surface, which method comprises the steps of:

cultivating cells on an electrode surface conductive to cell adhesion and growth which forms part of a device for subjecting adherent cells to an electrical field, wherein said device for subjecting adherent cells to an electrical field also has means provided for detecting he electrically induced polarization of an electroporation medium;
replacing the culturing medium with an electroporation medium having in suspension exogenous material desired to be entered into the cells;
applying an increasing electrical potential to said device under control of a modulating signal;
detecting the electrically induced polarization of the electro-poration medium used in said device;
discriminating a transmembrane flow of ions from cells when the cells porate
converting said discriminated transmembrane flow of ions into said modulating signal;
and
replacing the electroporation medium with culturing medium.

5 more elements

Dependencies are inferred from the claim text.

Method and device for cell cultivation on electrodes

Life of this patent

Method and device for cell cultivation on electrodes

Life of this patent

Abstract

Claims22