USE OF BONE MARROW CELLS FOR LONG TERM CULTURE OF PANCREATIC ISLET CELLS
Bone marrow cells have been demonstrated to be useful in the maintenance of pancreatic islet β cell viability, structure, and/or function in culture for a sustained period. Bone marrow cells were also found to promote β cell growth while reducing inflammatory cytokine release and reduce apoptosis. Moreover, islet cells co-cultured with bone marrow cells were shown to retain insulin response function and to function in an islet cell transplant in a mouse model of diabetes to restore normal insulin secretion. Cord blood cells and isolated peripheral CD34+ blood cells were unable to support β islet cell growth or increase survival.
1 . A method for culturing islet β cell comprising culturing an islet β cell with a bone marrow cell in vitro.
2 . The method of claim 1 , wherein the islet β cell is cultured for a sustained period.
3 . The method of claim 2 , wherein the sustained period is at least about 30 days, at least about 60 days, at least about 90 days or at least about 120 days.
4 - 6 . (canceled)
7 . The method of claim 1 , wherein culturing an islet β cell with a bone marrow cell increases islet β cell viability, improves islet β cell function, and/or improves islet β cell morphology as compared to an islet β cell cultured in the absence of a bone marrow cell.
8 . The method of claim 7 , wherein cell function is determined by a basal insulin secretion assay or a glucose induced insulin secretion assay.
9 . (canceled)
10 . The method of claim 7 , wherein increased islet β cell viability, improved islet β cell function, and/or improved islet β cell morphology are achieved for a sustained period.
11 . The method of claim 1 , wherein the method reduces apoptosis of an islet β cell in culture with a bone marrow cell as compared to an islet β cell cultured in the absence of a bone marrow cell.
12 . The method of claim 1 , wherein the method reduces cytokine release from an islet β cell in culture with a bone marrow cell as compared to an islet β cell cultured in the absence of a bone marrow cell.
13 . The method of claim 12 , wherein the cytokine is IL-1 β.
14 . The method of claim 1 , wherein the method increases the expression of at least one endocrine cell specific genes.
15 . The method of claim 14 , wherein the endocrine cell specific genes are selected from the group consisting of GCG (glucagon, α-cell gene), INS (insulin, β-cell gene), and SST (somatostatin, γ-cell gene); transcription factors for β cell pancreatic and duodenal homeobox1 (PDX1 or IPF1), Neurogenin 3 (NGN3), paired box gene 6 (PAX6), islet-1 (ISL1), v-maf musculoaponeurotic fibrosarcoma oncogene homolog A (MAFa), and Mist 1.
16 . A method for pancreatic islet β cell expansion in vitro comprising, culturing an islet β cell with a bone marrow cell.
17 . A method for increasing islet β cell viability, improved islet β cell function, and/or improved β cell morphology comprising culturing an islet β cell with a bone marrow cell in vitro.
18 - 24 . (canceled)
25 . A method of treating a subject in need of an islet β cell transplant comprising culturing an islet β cell with a bone marrow cell in vitro; and implanting the islet cell and the bone marrow cell into the individual in need of an islet β cell transplant.
26 - 30 . (canceled)
31 . A method of treating diabetes in a subject in need thereof comprising:
culturing an islet β cell with a bone marrow cell in vitro; and
implanting the cultured islet β cell into the subject;
thereby treating the subject for diabetes.
32 - 36 . (canceled)
37 . A method for implanting islet β cells in a subject comprising:
culturing an islet β cell with a bone marrow cell in vitro; and
implanting the cultured islet β cell into the subject.
38 - 41 . (canceled)
42 . A kit comprising a bone marrow cell and instructions for use in accordance with the method of claim 1 .