Three-dimensional ex vivo angiogenesis system
First Claim
1. A method for assaying the angiogenic potential of a particular tumor in a mammal;
- said method comprising the steps of;
(a) embedding a three-dimensional tissue sample in a matrix, wherein the tissue sample is taken from a particular tumor in a mammal;
wherein the tissue sample has at least one cut surface exposing blood vessels;
wherein the three-dimensional tissue sample comprises multiple layers of cells comprising blood vessels, supportive stromal elements, neural cells, and endothelial cells;
wherein the architecture of the tissue sample is substantially intact and has not been disrupted as compared to that of comparable tissue in vivo; and
wherein the three-dimensional tissue sample does not consist of an isolated artery or an isolated vein;
(b) supplying to the embedded tissue sample a medium that supports the growth of the tissue sample;
(c) incubating the embedded tissue sample in the medium for a time sufficient to allow any angiogenic vessels to grow into the matrix surrounding the tissue sample; and
(d) observing or measuring any angiogenic vessels that grow into the matrix surrounding the tissue sample;
whereby;
the growth or any angiogenic vessels into the matrix is a measure of the angiogenic potential of the particular tumor from which the tissue sample was taken.
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Abstract
An in vitro tissue angiogenesis and vasculogenesis system is disclosed that allows the outgrowth of microvessels from a three-dimensional tissue fragment implanted in a matrix. The matrix may, for example, be a fibrin- or collagen-based matrix fed by a growth medium, for example, a mixture of tissue culture medium, serum, or a layer of growth medium containing a defined mixture of growth factors. This system, which may be used with human or other mammalian or animal tissues, may be used in assaying tumor angiogenic potential, or in promoting angiogenesis in other tissues, e.g., promoting angiogenesis prior to transplantation of a tissue. The angiogenic potential of a tissue can be determined by measuring the growth of microvessels into the matrix. The three-dimensional structure of the tumor or other tissue is maintained in the matrix, including blood vessels. In another aspect, the method allows for the proliferation of a tissue specimen, thus increasing the mass of cells available for subsequent transplant; and the method also provides for the proliferation of blood vessels from the tissue mass, thus enhancing the chance of successful engraftment.
41 Citations
13 Claims
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1. A method for assaying the angiogenic potential of a particular tumor in a mammal;
- said method comprising the steps of;
(a) embedding a three-dimensional tissue sample in a matrix, wherein the tissue sample is taken from a particular tumor in a mammal;
wherein the tissue sample has at least one cut surface exposing blood vessels;
wherein the three-dimensional tissue sample comprises multiple layers of cells comprising blood vessels, supportive stromal elements, neural cells, and endothelial cells;
wherein the architecture of the tissue sample is substantially intact and has not been disrupted as compared to that of comparable tissue in vivo; and
wherein the three-dimensional tissue sample does not consist of an isolated artery or an isolated vein;
(b) supplying to the embedded tissue sample a medium that supports the growth of the tissue sample;
(c) incubating the embedded tissue sample in the medium for a time sufficient to allow any angiogenic vessels to grow into the matrix surrounding the tissue sample; and
(d) observing or measuring any angiogenic vessels that grow into the matrix surrounding the tissue sample;
whereby; the growth or any angiogenic vessels into the matrix is a measure of the angiogenic potential of the particular tumor from which the tissue sample was taken. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
- said method comprising the steps of;
Specification