Abstract: Implant materials having porosity are provided by the present invention. Methods for treating a synthetic or naturally occurring implant, such as a vascular graft, intended for implantation in a human patient are also provided comprising obtaining human microvascular fat tissue, homogenizing the fat tissue to provide a cellular slurry, and applying the cellular slurry onto said implant to provide at least 50% confluence of said cells on the surface and/or within the pores of the implant.

Abstract: Apparatus for depositing a cell product, such as endothelial cell product, in a graft and inserting the graft in a vessel is disclosed. The apparatus preferably comprises a tunneler tube that has a hollow portion for supporting the graft therein, apertures to permit the flow of cell product, and a pointed end cap attached to a distal end of the tunneler tube. The apparatus also has a handle connected to the graft and releaseably connected to a proximal end of the tunneler tube that provides an inlet for cell product and outlet in fluid communication with the lumen of the graft. During deposition, the cell product thus flows through the handle, into the graft and exits through the apertures. During insertion, the tunneler tube is manipulated by the handle to enter a vessel, and is then released from the handle and removed to accommodate anastomoses.

Abstract: Methods and apparatus for collecting and processing tissue to produce an endothelial cell product having a vessel for rinsing, draining, digesting and isolating tissue. The vessel has a rinsing and digesting chamber for containing tissue during processing. An inlet in the rinsing and digesting chamber allows entry of rinsing solution and tissue from a liposuction device. A waste chamber in fluid communication with the rinsing and digesting chamber preferably connects with a vacuum source. An isolation chamber is separated from the rinsing and digesting chamber by a screen. An ampule in fluid communication with the isolation chamber includes a pair of ports controlled by valve devices to be selectively in fluid communication with the isolation chamber. After processing, the ampule isolates a pellet of endothelial cells and the valve devices permit the pellet to be in fluid communication with the ports.

Abstract: The invention is an endothelial cell procurement and deposition kit for collecting fat from a patient, processing said fat to produce an endothelial cell deposition product, and depositing said product on the surface of a graft, all under sterile conditions established and maintained within the components of said kit comprised of: fat collection means for collecting subcutaneous fat from a patient; digestion means connectable to said fat collection means to maintain sterility during reception of said fat and for retaining said fat under sterile conditions during rinsing and digestion to produce a digested product; endothelial cell isolation means connectable to said digestion means for maintaining sterile conditions during reception of said digested product and for separating and isolating microvessel endothelial cells from said digested product to produce an endothelial cell product; cell deposition means connectable to said isolation means for maintaining sterile conditions during reception of said endothel

Abstract: The present invention provides a synthetic or naturally occurring implant, such as a vascular graft, for implantation into a human patient. Uncultured, microvascular endothelial cells, isolated from microvascular endothelial cell rich tissue are disposed on at least one surface of the implant to provide at least about 50% confluence of the cells on the surface of the implant prior to implantation. In a preferred embodiment, the microvascular endothelial cells are obtained from fat tissue. Because of the large number of fresh microvascular endothelial cells that may be obtained from such tissue, sufficient cells may be placed on the implant so that they attach to provide at least 50% confluent coverage of the implant surface prior to the time of implantation.

Abstract: Determination of effectiveness of microvascular endothelial cell seeding upon a vascular graft surface within the operating room environment would be desirable to maintain quality control in any clinical trial. A number of fluorescent dyes including mithramycin, Hoechst 33342, sulfofluorescein diacetate, Nile Red, rhodamine 123, and PKH26-GL were evaluated for their ability to fluorescently label uncultured microvascular endothilial cells on graft material and subsequently allow determination of seeded cell number and cell spreading. Rhodamine 123 and PKH26-GL produced the most desirable characteristics. The selected non-toxic fluorescent dyes allowed excellent cell visualization after a 30 minute incubation.

Abstract: A novel implantable prosthesis for implantation in human patients is disclosed which comprises a synthetic substrate and a Type IV/V collagen surface layer seeded with a confluent monolayer of autologous endothelial cells. In the preferred embodiment, a base layer of interstitial collagen is adhered to the substrate. The Type IV/V collagen surface layer is provided in a laminate comprising an intermediate layer of interstitial collagen which is covalently bound to the aforementioned base layer. A cross linking agent, such as glutaraldehyde, is utilized to covalently bind the interstitial collagen base layer to the substrate, and the collagen intermediate layer to the base layer. The cross linking agent is subsequently deactivated with a soluble peptide, such as lysine. Autologous endothelial cells, such as microvascular endothelial cells derived from adipose tissue, are then seeded at high densities on the pretreated graft material.

Abstract: Vigorous growth of endothelial cells from human blood vessels in vitro is achieved using a culture medium containing endothelial cell growth factor and heparin and/or dextran sulfate. Cell population doubling times of 13 to 21 hours for 60 to 85 populations doublings are obtained, and cloned human endothelial cell strains are established having similar proliferative capacities.

Abstract: The invention is an endothelial cell procurement and deposition kit for collecting fat from a patient, processing said fat to produce an endothelial cell deposition product, and depositing said product on the surface of a graft, all under sterile conditions established and maintained within the components of said kit comprised of: fat collection means for collecting subcutaneous fat from a patient; digestion means connectable to said fat collection means to maintain sterility during reception of said fat and for retaining said fat under sterile conditions during rinsing and digestion to produce a digested product; endothelial cell isolation means connectable to said digestion means for maintaining sterile conditions during reception of said digested product and for separating and isolating microvessel endothelial cells from said digested product to produce an endothelial cell product; cell deposition means connectable to said isolation means for maintaining sterile conditions during reception of said endothel

Abstract: Vigorous growth of endothelial cells from human umbilical vein endothelial cells in vitro is achieved using a culture medium containing endothelial cell growth factor and heparin and/or dextran sulfate. Cell population doubling times of 13 to 21 hours for 60 to 85 populations doublings are obtained, and cloned human endothelial cell strains are established having similar proliferative capacities.

Abstract: A method for treating the vascular passage of a patient, damaged by procedures such as an endarterectomy which denude portions of the vascular passages of their endothelial cell linings, is disclosed. In this method, endothelial cells are isolated from the patient's own microvessels, the flow of blood through the patient's damaged vascular passage is interrupted, the endothelial cells isolated from the patient's microvessels are applied to the surface of the denuded portion of the patient's vascular passage in a density sufficient to provide covereage of at least about 50% of said denuded portion, and interruption of blood flow through the vascular passage is maintained for a period of time sufficient to allow the sodded cells to form an attachment to the vascular lining sufficient to withstand the shear created by resumed blood flow through the vascular passage.

Abstract: The present invention provides a novel method of treating a synthetic or naturally occurring implant, such as a vascular graft, intended for implantation in a human patient, comprising obtaining human microvascular rich tissue from that patient; separating microvascular endothelial cells from that tissue; and placing said microvascular endothelial cells onto said implant to provide at least about 50% confluence of said cells on the surface of said implant to be treated. In the preferred embodiment, the microvascular rich tissue is perinephric fat, which is obtained from the donor and subjected to a digestive separation process to provide an abundant supply of microvascular endothelial cells. In the preferred embodiment, these microvascular endothelial cells are placed onto the implant by suspending them in a protein (plasma) containing physiologic saline solution, which is incubated on the graft until the endothelial cells either adhere to the graft surface or clot within the protein solution.

Abstract: A peritoneal dialysis apparatus comprising a plurality of components including a catheter, a fluid heater, a fluid weighing means and a control device. The components are interconnected by conduits so that fluid can flow from the heater to the peritoneal cavity by way of the catheter, and then from the peritoneal cavity to the weighing means. The weighing means measures the quantity of discharged fluid as a result of the introduction of the dialysis fluid to check that the dialysis fluid is causing fluid to drain from the peritoneal cavity. The flow of fluid between the components of the system is controlled by valves which are selectively opened and closed.