Insulin-producing bone marrow derived cells and methods of generating and using same

• US 20060216277A1
• Filed: 09/02/2004
• Published: 09/28/2006
• Est. Priority Date: 09/15/2003
• Status: Abandoned Application

• Current Assignee: Ramot at Tel Aviv University Limited (Tel Aviv University)
• Original Assignee: Ramot at Tel Aviv University Limited (Tel Aviv University)
• Inventors: Efrat, Shimon
• Application Number:
  US10/571,770
• Publication Number:
  US 20060216277A1
• Time in Patent Office: Days
• US Class Current: 424/93.210
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin
• CPC Class Codes: A61P 7/12 Antidiuretics, e.g. drugs for diabetes insipidus ADH A61P5/10
• CPC Class Codes: C12N 2506/1353 from bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2510/02 Cells for production
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0676 Pancreatic cells

Abstract

Insulin-producing bone marrow derived stem cells, and methods of generating and using same to reduce blood glucose levels in individuals.

Claims

1-74. -74. (canceled)

75. A method of generating insulin-producing cells comprising:

(a) isolating and optionally culturing adult tissue stem cells to thereby obtain an adult tissue stem cell culture; and

(b) expressing exogenous Pdx1 in cells of said adult tissue stem cell culture to thereby obtain insulin-producing cells.

76. The method of claim 75, further comprising a step of immortalizing said cells of said adult tissue stem cell culture.

77. The method of claim 76, wherein said immortalizing is effected by expressing in said cells of said adult tissue stem cell culture a telomerase.

78. The method of claim 75, wherein said adult tissue stem cells are selected from the group consisting of adipose tissue stem cells, skin stem cells, kidney stem cells, liver stem cells, prostate stem cells, pancreas stem cells, intestine stem cells and bone marrow stem cells.

79. The method of claim 78, wherein said adult tissue stem cells are bone marrow stem cells.

80. The method of claim 75, wherein said culturing is effected under culturing conditions selected suitable for expansion of mesenchymal stem cells of said bone marrow stem cells.

81. The method of claim 75, wherein said expressing in said cells said exogenous Pdx1 is effected by transfecting said cells with an expression vector including a polynucleotide encoding said Pdx1 positioned under the transcriptional control of a mammalian promoter.

82. A cell culture comprising adult tissue stem cells genetically modified to express Pdx 1.

83. The cell culture of claim 82, wherein said adult tissue stem cells are selected from the group consisting of adipose tissue stem cells, skin stem cells, kidney stem cells, liver stem cells, prostate stem cells, pancreas stem cells, intestine stem cells and bone marrow stem cells.

84. The cell culture of claim 82, wherein said adult tissue stem cells are bone marrow stem cells.

85. The cell culture of claim 82, wherein said cells are capable of producing insulin.

86. The cell culture of claim 82, wherein said cells are capable of expressing pancreatic beta cell genes.

87. The cell culture of claim 82, wherein said adult tissue stem cells stem cells are immortalized.

88. The cell culture of claim 87, wherein said adult tissue stem cells stem cells are genetically modified to express a telomerase.

89. A method of reducing blood glucose levels in an individual comprising administering Pdx1-expressing adult tissue stem cells to the individual thereby reducing blood glucose levels in the individual.

90. The method of claim 89, wherein said Pdx1-expressing bone marrow stem cells are prepared by:

(a) isolating and optionally culturing adult tissue stem cells to thereby obtain an adult tissue stem cell culture; and

(b) expressing exogenous Pdx1 in cells of said adult tissue stem cell culture to thereby obtain Pdx1-expressing bone marrow stem cells.

91. The method of claim 90, further comprising a step of immortalizing said cells of said adult tissue stem cell culture.

92. The method of claim 91, wherein said immortalizing is effected by expressing in said cells of said adult tissue stem cell culture a telomerase.

93. The method of claim 89, wherein said adult tissue stem cells are selected from the group consisting of adipose tissue stem cells, skin stem cells, kidney stem cells, liver stem cells, prostate stem cells, pancreas stem cells, intestine stem cells and bone marrow stem cells.

94. The method of claim 89, wherein said adult tissue stem cells are bone marrow stem cells.

95. The method of claim 94, wherein said culturing is effected under culturing conditions selected suitable for expansion of mesenchymal stem cells of said bone marrow stem cells.

96. The method of claim 90, wherein said expressing in said cells said exogenous Pdx1 is effected by transfecting said cells with an expression vector including a polynucleotide encoding said Pdx1 positioned under the transcriptional control of a mammalian promoter.

97. A method of treating an individual having a disorder requiring β

-cell replacement, comprising administering Pdx1-expressing adult tissue stem cells to the individual thereby treating the individual.

98. The method of claim 97, wherein said Pdx1-expressing adult tissue stem cells are prepared by:

(a) isolating and optionally culturing adult tissue stem cells to thereby obtain an adult tissue stem cell culture; and

(b) expressing exogenous Pdx1 in cells of said adult tissue stem cell culture to thereby obtain Pdx 1-expressing adult tissue stem cells.

99. The method of claim 98, further comprising a step of immortalizing said cells of said adult tissue stem cell culture.

100. The method of claim 99, wherein said immortalizing is effected by expressing in said cells of said adult tissue stem cell culture a telomerase.

101. The method of claim 97, wherein said adult tissue stem cells are selected from the group consisting of adipose tissue stem cells, skin stem cells, kidney stem cells, liver stem cells, prostate stem cells, pancreas stem cells, intestine stem cells and bone marrow stem cells.

102. The method of claim 97, wherein said adult tissue stem cells are bone marrow stem cells.

103. The method of claim 102, wherein said culturing is effected under culturing conditions selected suitable for expansion of mesenchymal stem cells of said bone marrow stem cells.

104. The method of claim 98, wherein said expressing in said cells said exogenous Pdx1 is effected by transfecting said cells with an expression vector including a polynucleotide encoding said Pdx1 positioned under the transcriptional control of a mammalian promoter.