Mesenchymal precursor cell

• US 20050281790A1
• Filed: 06/29/2005
• Published: 12/22/2005
• Est. Priority Date: 07/07/1999
• Status: Active Grant

• Current Assignee: Mesoblast, Inc. (Mesoblast Ltd.)
• Original Assignee: Medvet Science Pty. Ltd.
• Inventors: Gronthos, Stan, Zannettino, Andrew, Simmons, Paul
• Granted Patent: US 8,158,118 B2
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: C12N 2500/42 Organic phosphate, e.g. beta glycerophosphate
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

A method of enriching mesenchymal precursor sells including the step of enriching for cells based on at least two markers. The markers may be either i) the presence of markers specific for mesenchymal precursor cells, ii) the absence of markers specific for differentiated mesenchymal cells, or iii) expression levels of markers specific for mesenchymal precursor cells. The method may include a first solid phase sorting step utilizing MACS recognizing expression of the antigen to the STRO-1 Mab, followed by a second sorting step utilizing two color FACS to screen for the presence of high level STRO-1 antigen expression as well as the expression of VCAM-1.

Claims

1-51. -51. (canceled)

52. An enriched population of mesenchymal precursor cells obtained by selecting for cells which are positive for one or more markers selected from the group consisting of STRO-1^bright, VCAM-1^bright, THY-1^bright, CD146^bright and STRO-2^bright or any combination thereof.

53. An enriched cell population as in claim 52 wherein the STRO-1^bright cells carry a high copy number of an antigen identified by STRO-1.

54. An enriched cell population as in claim 52 wherein the VCAM-1^bright cells carry a high copy number of an antigen identified by VCAM-1.

55. An enriched cell population as in claim 52 wherein the THY-₁^bright cells carry a high copy number of an antigen identified by THY-1.

56. An enriched cell population as in claim 52 wherein the CD146^bright cells carry a high copy number of an antigen identified by CD146.

57. An enriched cell population as in claim 52 wherein the STRO-2^bright cells carry a high copy number of an antigen identified by STRO-2.

67. An enriched population of cells as in either of claim 52 or claim 58 wherein a proportion of the cells are capable of differentiation into at least two committed cell types selected from the group including adipose, areolar, osseous, cartilaginous, elastic and fibrous connective tissue.

68. An enriched population of cells as in either of claim 52 or claim 58 wherein the enriched population is suitable for seeding onto a vehicle for implantation to assist in bone growth.

69. An enriched population of cells as in either of claim 52 or claim 58 wherein the enriched population has an exogenous nucleic acid transformed in to it so that the population may be introduced into the body of a patient to treat a disease or condition.

70. An enriched population of cells as in either of claim 52 or claim 58 wherein the enriched population has an exogenous nucleic acid that expresses a therapeutic agent transformed in to it so that the population may be introduced into the body of a patient to release the therapeutic agent.

71. An enriched population of cells as in either of claim 52 or claim 58 wherein the enriched population is used to augment bone marrow transplantation.

72. A composition including the enriched population of claim 52.

74. A composition as in either of claim 72 or claim 73 wherein the composition is preadsorbed onto ceramic vehicles that are precoated with fibronectin and are suitable for implantation to augment bone marrow transplantation.

75. A composition as in either of claim 72 or claim 73 wherein the composition is suitable for use in augmenting bone marrow transplantation.

76. A composition as in claim 75 wherein the composition also includes haemopoietic cells.

77. A composition as in either of claim 72 or claim 73 wherein the population has an exogenous nucleic acid transformed in to it so that the composition may be introduced into the body of a patient to treat a disease or condition.

78. A composition as in either of claim 72 or claim 73 wherein the population has an exogenous nucleic acid that expresses a therapeutic agent transformed in to it so that the composition may be introduced into the body of a patient to release the therapeutic agent.

58. An enriched cell population wherein at least 1% of the cells are STRO-1^bright.

59. An enriched cell population as in claim 58 wherein at least 1.5% of the cells are STRO-1^bright.

60. An enriched cell population as in claim 58 wherein at least 2% of the cells are STRO-1^bright.

61. An enriched cell population as in claim 58 wherein at least 5% of the cells are STRO-1^bright.

62. An enriched cell population as in claim 58 wherein at least 10% of the cells are STRO-1^bright.

63. An enriched cell population as in claim 58 wherein at least 40% of the cells are STRO-1^bright.

64. An enriched cell population as in claim 58 wherein the STRO-1^bright cells carry a high copy number of an antigen identified by STRO-1.

65. An enriched cell population as in claim 58 wherein the STRO-1^bright cells carry at least one additional marker selected from a group of surface markers specific for mesenchymal precursor cells consisting of LFA-3, THY-1, antigen identified by STRO-1, VCAM-1, ICAM-1, PECAM-1, P-selectin, L-selectin, CD49b/CD29, CD49c/CD29, CD49d/CD29, CD29, CD18, CD61, 6-19, thrombomodulin, CD10, CD13, integrin beta, STRO-2, CD146, and SCF or combinations thereof.

66. An enriched cell population as in claim 58 wherein the STRO- 1^bright cells are negative for at least one marker selected from the group consisting of CBFA-1, collagen type II, PPARγ

2, and glycophorin A.

73. A composition including the enriched population of claim 58.