Method For Production Of Large Numbers Of Cartilage Cells With Phenotype Retention
A method for production of large numbers of cartilage cells with phenotype retention intended for treatment of articular cartilage lesions or preparing viable cartilage tissue by propagation of chondrocytes from cartilage explants, human or animal, with the retention of phenotypes of the cartilage cells, in which cells are chondrocytes retaining morphologic attributes of the same or chondrocyte progenitor cells. The method includes culturing the cells in which cultured cells are organized into mature hyaline cartilage on the surface of biologic structures such as cancellous or cortical bone lamina. The method for culturing chondrocytes to produce three dimensional cellular structures consisting of cartilage cells and cell produced extracellular cartilage matrix.
1 . A method for production of large numbers of cartilage cells comprises the step of:
preparing viable cartilage tissue by propagation of chondrocytes from cartilage explants, human or animal, with the retention of phenotypes of the cartilage cells.
2 . The method according to claim 1 in which cells are chondrocytes retaining morphologic attributes of the same or chondrocyte progenitor cells.
3 . The method according to claim 1 in which cultured cells are organized into mature hyaline cartilage on the surface of biologic structures such as cancellous or cortical bone lamina.
4 . The method according to claim 1 for culturing chondrocytes to produce three dimensional cellular structure consisting of cartilage cells and cell produced extracellular cartilage matrix.
5 . The method according to claim 1 whereby biologic material comprising hyaline cartilage tissue is propagated on the surface of thin cancellous bone which is undecalcified, and wherein the said cancellous bone lamina is from 0.5 to 2.0 mm in thickness.
6 . The method according to claim 1 whereby cancellous bone lamina, 0.5 to 2.0 mm thick is decalcified to render it flexible.
7 . The method according to claim 1 whereby cancellous bone lamina is either demineralized or undemineralized and is thicker than 2.0 mm.
8 . The method according to claim 5 whereby newly proliferating cartilage can be transplanted into a chondral defect together with the underlying substrate without expanding the cells.
9 . The method according to claim 6 whereby newly proliferating cartilage can be transplanted into a chondral defect together with the underlying substrate without expanding the cells.
10 . The method according to claim 7 whereby newly proliferating cartilage can be transplanted into a chondral defect together with the underlying substrate without expanding the cells.
11 . The method according to claim 1 whereby a substrate on which chondrocytes proliferate from explants is a lamina of undecalcified cortical bone 0.5 to 2.0 mm thick with multiple, geometrically placed perforations.
12 . The method according to claim 1 whereby the lamina on which explants are placed is decalcified cortical bone plate 0.5 to 2.0 mm thick with multiple, geometrically placed perforations.
13 . The method according to claim 1 whereby all osseous substrates on which chondrocytes have proliferated from cartilage explants are cryopreserved for further transplantation.
14 . The method according to claim 1 whereby cartilage explants are cultivated on a cellular dermis, pericardium, dura mater and fascia with or without perforations.
15 . The method according to claim 1 whereby chondrocytes proliferated on soft membranes with or without perforations are cryopreserved for future transplantation.
16 . The method according to claim 1 whereby cartilage explants are placed on Gelfoam®, Surgicel®, Gelatin USP, and or similar materials which can be optionally supplemented with thrombin or ligrin.
17 . The method according to claim 1 whereby cartilage explants are grown on membranes of collodion, polyvinylpyrrolidone, hydroxyethyl starch or an equivalent thereof.
18 . The method according to claim 1 whereby cartilage cells together with or without cartilage explants are harvested mechanically, enzymatically or physically from the substrates on which they are grown, frozen and either freeze-dried or dehydrated under hypothermia and micronized for implantation into articular defects.
19 . The method according to claim 18 wherein cells and explants can be frozen with substrates and then later scraped with a blade for further processing.