Advanced Tissue Sciences, Inc.

Noushin Dunkelman

Alvin Edward Peterson

Gail K Naughton

Joan Zeltinger

Lee K Landeen

Brian A Naughton

Stephen V Kemmerrer

Mark A Applegate

James H Flatt

Anthony F Purchio

John W Bennett

Scott Chesla

Jane Willoughby

Jason Gee

Michael Zimber

Ronda Elizabeth Schreiber

Dror Seliktar

An apparatus and method for sterilizing, seeding, culturing, storing, shipping, and testing three-dimensional tissue constructs is disclosed. The apparatus includes a fluid reservoir, a pump, at least one treatment chamber, and a means for controlling media flow characteristics around a tissue construct disposed within the treatment chamber, and for controlling movement of the construct itself, so as to simulate a variety of physiologic conditions. One exemplary embodiment of the invention includes a means for applying an axial stress to the construct. Applying an axial stress to the construct during seeding and culturing results in a tissue-engineered construct with cells and their fibers oriented in a manner which is more likely to possess long term dimensional stability and the patency of native vessels with normal physiologic function.

Apparatus and method for simulating in vivo conditions while seeding and

Mammalian cells capable of producing cartilage are cultured under a shear flow stress of about 1 to about 100 dynes/cm.sup.2 to produce artificial cartilage for surgical transplantation to replace damaged or missing cartilage. Shear flow stressed cells display enhanced maintenance of chondrocyte phenotype and produce an extracellular matrix containing an enhanced ratio of type II collagen to type I collagen. The cells may be chondrocytes, chondrocyte stem cells or cells such as myoblasts or fibroblasts that transdifferentiate into chondrocytes. A bioreactor is used containing a growth chamber having a substrate on which the cells capable of producing cartilage are attached, and means for applying relative movement between a liquid culture medium and the substrate to provide the shear flow stress. The substrate may be a scaffold, or a nonporous surface such as the surface of a rotatable drum or disc, or the surface of a static plate that supports cell growth in a monolayer. Cartilage is produced by covering the substrate with a liquid growth medium, inoculating into the medium cells capable of producing cartilage, allowing the cells to attach to the substrate, applying and maintaining relative movement between the growth medium and the cells to provide the shear flow stress to produce shear flow stressed cells, and culturing the shear flow stressed cells to produce cartilage.

Application of shear flow stress to chondrocytes or chondrocyte stem

The present invention relates to a method of stimulating the proliferation and appropriate cell maturation of a variety of different cells and tissues in three-dimensional cultures in vitro using TGF-.beta. in the culture medium. In accordance with the invention, stromal cells, including, but not limited to, chondrocytes, chondrocyte-progenitors, fibroblasts, fibroblast-like cells, umbilical cord cells or bone marrow cells from umbilical cord blood are inoculated and grown on a three-dimensional framework in the presence of TGF-.beta.. Stromal cells may also include other cells found in loose connective tissue such as endothelial cells, macrophages/monocytes, adipocytes, pericytes, reticular cells found in bone marrow stroma, etc. The stromal cells and connective tissue proteins naturally secreted by the stromal cells attach to and substantially envelope the framework composed of a biocompatible non-living material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells. The living stromal tissue so formed provides the support, growth factors, and regulatory factors necessary to sustain long-term active proliferation of cells in culture and/or cultures implanted in vivo. When grown in this three-dimensional system, the proliferating cells mature and segregate properly to form components of adult tissues analogous to counterparts in vivo.

Three-dimensional cartilage cultures

An apparatus and method for sterilizing, seeding, culturing, storing, shipping, and testing vascular grafts is disclosed. Specifically, the present invention relates to an apparatus and method for seeding and culturing vascular grafts with human cells. The apparatus includes a fluid reservoir, a pump, an alternating pressure source, and at least one treatment chamber. By alternating pressure to a support structure within the treatment chamber upon which a vascular graft scaffold is positioned, a varying radial stress is placed on the scaffold. In an alternative embodiment, fluid is pumped directly through the vascular graft subjecting the vascular graft to radial and shear stresses. Applying shear and/or radial stresses to the vascular graft during seeding and culturing simulates physiological conditions.

Apparatus and method for sterilizing, seeding, culturing, storing,

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San Diego, CA, United States of America

Noushin Dunkelman