Cedars-sinai Medical Center

Pacfab, Inc.

Jacek Rozga

Achilles A Demetriou

Andreas Kamlot

Eugenio Morsiani

Disclosed is a universally-adaptable cell preconditioning and storage medium that can be used for the effective cryopreservation of cells at temperatures less than 4&deg; C. The aqueous medium contains adenosine, a calcium channel blocker, and a nutrient-rich matrix that has a sufficient amount of cell metabolites to sustain the metabolic needs of the harvested cells while incubating the cells for a period of at least 10 minutes, without producing detectable levels of lactate or substantially depleting the metabolic substrates of the cell.

Cell preconditioning and cryopresevation medium

An artificial gut ( ) comprises a first hollow fiber ( ) having an inner surface ( ) that is lined with at least a portion of a layer of one or more of a plurality of biological components ( ) that typically line a mammalian gut. Further, a second hollow fiber ( ) is adjacent the first hollow fiber. The layer of one or more of the plurality of biological components can consist essentially of enterocytes. The enterocyte-lined inner surface is perfused with a feeding solution containing nutrients, and the enterocytes absorb, process, and transport the nutrients across the wall of the first hollow fiber. The nutrients eventually diffuse into the second hollow fiber. A perfusate selected from the group consisting of culture medium, blood, and plasma, can perfuse an inner surface of the second hollow fiber, whereby the perfusate is nourished by the nutrients.

Artificial gut

A bioreactor (,) has arranged within its internal chamber (,) a bundle of liquid-impermeable hollow tubes (,), which are used to freeze or vitrify a biologically active material seeded within the internal chamber. When the bioreactor is ready for use, the biologically active material may be thawed by perfusing the liquid-impermeable hollow tubes with a heated solution or heated vapor.

Bioreactor and related method

A bioreactor (,) comprises an elastic housing (,) and a plurality of hollow, porous fibers (,) disposed within the housing. The lumens of the fibers define an intrafiber compartment, and the outer surfaces of the fibers and the housing define an extrafiber compartment. A variable flow device (,) varies the flow of the perfusate through the intrafiber compartment. The variable flow device can include a flow restrictor (,) that variably restricts the discharge of a perfusate (,) from the intrafiber compartment or a pump for increasing the flow of the perfusate into the intrafiber compartment. The elastic housing can include a wall (,) with perforations (,) extending therethrough and an elastic membrane (,) tightly surrounding the wall or a wall with at least one expansion port extending therethrough and at least one extrafiber space expander,coupled to a one of the at least one expansion port.

A method is provided for preconditioning and cryopreservation of cells harvested from a donor. Cells are suspended in a cell conditioning and cryopreservation medium containing a cryopreservative such as Dimethyl Sulfoxide (DMSO) and the suspension is incubated for a period of at least ten minutes and the cells are frozen. The medium includes adenosine, a calcium channel blocker, and a cell nutrient matrix comprising a sufficient amount of cell nutrients to sustain the metabolic needs of the cells during incubation without producing detectable levels of lactate or substantially depleting the nutrient substrates to maintain viability of the harvested cells. In some embodiments, a cryopreservative is added step-wise before the cell suspension is frozen and removed step-wise after the cell suspension is thawed.

Method for conditioning and cryopreserving cells

An artificial gut (100) comprises a first hollow fiber (203) having an inner surface (215) that is lined with at least a portion of a layer of one or more of a plurality of biological components (205) that typically line a mammalian gut. Further, a second hollow fiber (201) is adjacent the first hollow fiber. The layer of one or more of the plurality of biological components can consist essentially of enterocytes. The enterocyte-lined inner surface is perfused with a feeding solution containing nutrients, and the enterocytes absorb, process, and transport the nutrients across the wall of the first hollow fiber. The nutrients eventually diffuse into the second hollow fiber. A perfusate selected from the group consisting of culture medium, blood, and plasma, can perfuse an inner surface of the second hollow fiber, whereby the perfusate is nourished by the nutrients.

The invention resides in apparatus and methods for isolating and collecting cells from donor tissue for later use in medical procedures. The apparatus includes a sealed enclosure, at least one screening material inside the enclosure, an inlet port configured to conduct a fluid through the screening material, and an outlet port for receiving the fluid from the screening material and conducting the fluid out of the enclosure. A material to be screened is placed inside or conducted into the enclosure. Screened material is entrained in the fluid. The fluid carries bits of the material through the screening material and out of the enclosure for later separation from the fluid. Preferred embodiments include a series of screening materials of decreasing pore size, usually in the range from ten to one thousand micrometers in diameter. A particularly preferred embodiment includes three distinct screening materials having pore sizes of 400 micrometers, 280 micrometers, and 100 micrometers, listed in the order that the fluid passes through the screening materials. The invention is used in the collection and isolation of porcine hepatocytes for later use in medical procedures. The sealed enclosure is constructed of a series of individual segments held together by conveniently removable fasteners--bolts and wing nuts. A mechanical shaker agitates the enclosure as the fluid flows over and through the material being screened.

System for hepatocyte cell isolation and collection

Methods for cell isolation and collection

An implantable catheter comprises a flexible elongated catheter body having an inflatable balloon positioned near its distal end. At least one distal delivery port is located distal of the balloon and at least one proximal delivery port is located proximal of the balloon. The balloon and the distal and proximal delivery ports are connected in fluid communication with individual lumens. The lumen connected to the balloon is connected to a balloon inflation port at the proximal end of the catheter. The lumens connected to the distal and proximal delivery ports are connected to distal and proximal injection ports. Preferably, the balloon inflation port and the distal and proximal injection ports are all built into an integrated port assembly adapted for implantation under the skin of a medical patient. In use, the distal end of the catheter is inserted and fed into a passageway such as a blood vessel until the balloon and delivery ports lie within a region of interest. The port assembly is then implanted under the patient's skin. Once in place, the balloon can be inflated to occlude the flow of fluid through the passageway, and therapeutic agents can be delivered either distal or proximal of the balloon by injection into the appropriate injection port. It is believed that the catheter is particularly suited to the delivery of hepatocytes and an anticoagulant during a course of cell therapy directed to a patient's liver.

Growing community of inventors

Westlake Village, CA, United States of America

Jacek Rozga