Method for treating connective tissue disorders

Claim 1

A method for repairing connective tissue damage comprising: a) providing culturally expanded purified human marrow-derived mesenchymal cells which have been isolated from a bone marrow specimen by adding the bone marrow specimen to a medium which contains factors which stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for selective adherence of only the mesenchymal stem cells to a substrate surface; and, b) applying the culturally expanded purified human marrow-derived mesenchymal cells to an area of connective tissue damage under conditions suitable for differentiating the cells into connective tissue cells necessary for repair.

Claim 2

A method for repairing connective tissue damage comprising: a) providing culturally expanded purified human marrow-derived mesenchymal cells which have been isolated from a bone marrow specimen by adding the bone marrow specimen to a medium which contains factors which stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for selective adherence of only the mesenchymal stem cells to a substrate surface; and, b) applying the culturally expanded purified human marrow-derived mesenchymal cells to an area of connective tissue damage under conditions suitable for differentiating the cells into connective tissue cells necessary for repair.

Claim 6

A method for enhancing implantation of a prosthetic device in connective tissue comprising: a) providing culturally expanded purified human marrow-derived mesenchymal cells which have been isolated from a bone marrow specimen by adding the bone marrow specimen to a medium which contains factors which stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for selective adherence of only the mesenchymal stem cells to a substrate surface; b) adhering said culturally expanded purified human marrow-derived mesenchymal cells on to a prosthetic device; and, c) implanting the prosthetic device containing the culturally expanded purified marrow-derived mesenchymal cells under conditions suitable for differentiating the cells into connective tissue.

Claim 7

A method for enhancing implantation of a prosthetic device in connective tissue comprising: a) providing culturally expanded purified human marrow-derived mesenchymal cells which have been isolated from a bone marrow specimen by adding the bone marrow specimen to a medium which contains factors which stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for selective adherence of only the mesenchymal stem cells to a substrate surface; b) adhering said culturally expanded purified human marrow-derived mesenchymal cells on to a prosthetic device; and, c) implanting the prosthetic device containing the culturally expanded purified marrow-derived mesenchymal cells under conditions suitable for differentiating the cells into connective tissue.

Claim 9

A method for repairing connective tissue damage comprising the steps of: a) providing a bone marrow specimen containing human mesenchymal stem cells and bone pieces; b) adding the bone marrow specimen to a medium which contains factors which stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for the selective adherence of only the mesenchymal stem cells to a substrate surface; c) separating the bone pieces from the bone marrow medium mixture; d) dissociating the marrow cells into single cells; e) culturing the dissociated marrow cells in the bone marrow medium mixture; f) separating the non-adherent matter from the substrate surface thereby producing isolated culturally expanded mesenchymal stem cells; and, g) applying the culturally expanded purified mesenchymal stem cells to the area of connective tissue damage under conditions suitable for differentiating the cells into the type of connective tissue cells necessary for repair.

Claim 10

A method for repairing connective tissue damage comprising the steps of: a) providing a bone marrow specimen containing human mesenchymal stem cells and bone pieces; b) adding the bone marrow specimen to a medium which contains factors which stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for the selective adherence of only the mesenchymal stem cells to a substrate surface; c) separating the bone pieces from the bone marrow medium mixture; d) dissociating the marrow cells into single cells; e) culturing the dissociated marrow cells in the bone marrow medium mixture; f) separating the non-adherent matter from the substrate surface thereby producing isolated culturally expanded mesenchymal stem cells; and, g) applying the culturally expanded purified mesenchymal stem cells to the area of connective tissue damage under conditions suitable for differentiating the cells into the type of connective tissue cells necessary for repair.

Claim 16

A method for enhancing implantation of a prosthetic device in a connective tissue comprising: a) providing a bone marrow specimen containing human mesenchymal stem cells and bone pieces; b) adding the bone marrow specimen to a medium which contains factors which stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for selective adherence of only the mesenchymal stem cells to a substrate surface; c) separating the bone pieces from the bone marrow medium mixture; d) dissociating the marrow cells into single cells; e) culturing the dissociated marrow cells in the bone marrow medium mixture; f) separating non-adherent matter from the substrate surface thereby producing isolated culturally expanded mesenchymal stem cells; g) adhering said culturally expanded purified mesenchymal stem cells on to a prosthetic device; and, h) implanting the prosthetic device containing the culturally expanded purified mesenchymal stem cells under conditions suitable for differentiating the cells into skeletal tissue.

Claim 17

A method for enhancing implantation of a prosthetic device in a connective tissue comprising: a) providing a bone marrow specimen containing human mesenchymal stem cells and bone pieces; b) adding the bone marrow specimen to a medium which contains factors which stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for selective adherence of only the mesenchymal stem cells to a substrate surface; c) separating the bone pieces from the bone marrow medium mixture; d) dissociating the marrow cells into single cells; e) culturing the dissociated marrow cells in the bone marrow medium mixture; f) separating non-adherent matter from the substrate surface thereby producing isolated culturally expanded mesenchymal stem cells; g) adhering said culturally expanded purified mesenchymal stem cells on to a prosthetic device; and, h) implanting the prosthetic device containing the culturally expanded purified mesenchymal stem cells under conditions suitable for differentiating the cells into skeletal tissue.

Claim 21

A method for generating a hematopoietic stem cell reserve comprising: a) providing human marrow-derived mesenchymal stem cells that have been isolated, purified and culturally expanded; b) applying the isolated, purified and culturally expanded marrow-derived human mesenchymal stem cells to a porous carrier; c) implanting the porous carrier containing the mesenchymal stem cells into a host patient; d) allowing for a sufficient period of time for inducement of hematopoietic stem cells present in the host patient into the porous carrier; and e) harvesting the hematopoietic stem cells present in the porous carrier.

Claim 22

A method for generating a hematopoietic stem cell reserve comprising: a) providing human marrow-derived mesenchymal stem cells that have been isolated, purified and culturally expanded; b) applying the isolated, purified and culturally expanded marrow-derived human mesenchymal stem cells to a porous carrier; c) implanting the porous carrier containing the mesenchymal stem cells into a host patient; d) allowing for a sufficient period of time for inducement of hematopoietic stem cells present in the host patient into the porous carrier; and e) harvesting the hematopoietic stem cells present in the porous carrier.

Claim 24

A method for generating a human hematopoietic stem cell reserve comprising: a) providing a bone marrow specimen containing human marrow-derived mesenchymal stem cells and bone pieces; b) adding the bone marrow specimen to a medium which contains factors which stimulate the human marrow-derived mesenchymal stem cells to grow without differentiation and allows, when cultured, for selective adherence of only the marrow-derived human mesenchymal stem cells to a substrate surface; c) separating the bone pieces from the bone marrow medium mixture; d) dissociating the marrow cells into single cells; e) culturing the dissociated marrow cells in the bone marrow medium mixture; f) separating non-adherent matter from the substrate surface, thereby producing isolated culturally expanded human marrow-derived mesenchymal stem cells; g) applying the culturally expanded human marrow-derived mesenchymal stem cells to a porous carrier; h) implanting the porous carrier containing the culturally expanded marrow-derived mesenchymal cells into a host patient; i) allowing for a sufficient period of time for inducement of hematopoietic stem cells present in the host patient into the porous carrier; and, j) harvesting the hematopoietic stem cells collected by the porous carrier.

Claim 25

A method for generating a human hematopoietic stem cell reserve comprising: a) providing a bone marrow specimen containing human marrow-derived mesenchymal stem cells and bone pieces; b) adding the bone marrow specimen to a medium which contains factors which stimulate the human marrow-derived mesenchymal stem cells to grow without differentiation and allows, when cultured, for selective adherence of only the marrow-derived human mesenchymal stem cells to a substrate surface; c) separating the bone pieces from the bone marrow medium mixture; d) dissociating the marrow cells into single cells; e) culturing the dissociated marrow cells in the bone marrow medium mixture; f) separating non-adherent matter from the substrate surface, thereby producing isolated culturally expanded human marrow-derived mesenchymal stem cells; g) applying the culturally expanded human marrow-derived mesenchymal stem cells to a porous carrier; h) implanting the porous carrier containing the culturally expanded marrow-derived mesenchymal cells into a host patient; i) allowing for a sufficient period of time for inducement of hematopoietic stem cells present in the host patient into the porous carrier; and, j) harvesting the hematopoietic stem cells collected by the porous carrier.

Claim 29

A method for repairing connective tissue damage comprising: a) providing a bone marrow specimen containing human mesenchymal stem cells; b) adding the bone marrow specimen to a medium thereby producing a bone marrow specimen-medium mixture, wherein said medium contains factors that stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for selective adherence of only the mesenchymal stem cells to a substrate surface; c) adding the bone marrow specimen-medium mixture to a density gradient which separates cells into low, medium and high density cell fractions based on differences in density; d) removing the low density cell fraction from the density gradient; e) adding the low density cell fraction to the medium used in step (b) to produce a low density cell fraction-medium mixture; f) culturing the low density cell fraction-medium mixture, thereby selectively adhering only the mesenchymal stem cells to the substrate surface; g) removing any non-adherent matter from substrate surface; h) removing the remaining adherent mesenchymal stem cells from the substrate surface with a releasing agent, thereby allowing for the isolated mesenchymal stem cells to be recovered; and, i) applying an culturally expanded purified human marrow-derived mesenchymal cells to the area of connective tissue damage under conditions suitable for differentiating the cells into connective tissue necessary for repair.

Claim 30

A method for repairing connective tissue damage comprising: a) providing a bone marrow specimen containing human mesenchymal stem cells; b) adding the bone marrow specimen to a medium thereby producing a bone marrow specimen-medium mixture, wherein said medium contains factors that stimulate mesenchymal stem cell growth without differentiation and allows, when cultured, for selective adherence of only the mesenchymal stem cells to a substrate surface; c) adding the bone marrow specimen-medium mixture to a density gradient which separates cells into low, medium and high density cell fractions based on differences in density; d) removing the low density cell fraction from the density gradient; e) adding the low density cell fraction to the medium used in step (b) to produce a low density cell fraction-medium mixture; f) culturing the low density cell fraction-medium mixture, thereby selectively adhering only the mesenchymal stem cells to the substrate surface; g) removing any non-adherent matter from substrate surface; h) removing the remaining adherent mesenchymal stem cells from the substrate surface with a releasing agent, thereby allowing for the isolated mesenchymal stem cells to be recovered; and, i) applying an culturally expanded purified human marrow-derived mesenchymal cells to the area of connective tissue damage under conditions suitable for differentiating the cells into connective tissue necessary for repair.