METHODS AND COMPOSITIONS RELATING TO PROGENITOR CELLS

• US 20100291219A1
• Filed: 06/20/2008
• Published: 11/18/2010
• Est. Priority Date: 06/21/2007
• Status: Abandoned Application

• Current Assignee: Brigham and Women's Hospital Incorporated, Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Sarkar, Debanjan, Karp, Jeffrey M., Graneli, Cecilia, Langer, Robert S.
• Application Number:
  US12/665,510
• Publication Number:
  US 20100291219A1
• Time in Patent Office: Days
• US Class Current: 424/489
• CPC Class Codes: A61F 2002/2817 Bone stimulation by chemical reactions or by osteogenic or biological products for enhancing ossification, e.g. by bone morphogenetic or morphogenic proteins [BMP] or by transforming growth factors [TGF]
• CPC Class Codes: A61K 35/32 Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12N 2533/56 Fibrin; Thrombin
• CPC Class Codes: C12N 5/0075 using microcarriers
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

The invention provides compositions and methods for harvesting and enriching cells such as connective tissue progenitors cells using stirred bioreactors, and in some embodiments fibrin microcarriers.

Claims

1. A method for isolating cells from a sample comprising

contacting a sample with a plurality of microcarriers in solution under conditions and for a time sufficient for cells in the sample to bind to the microcarriers, and

harvesting from the solution microcarriers bound to cells,

wherein the sample is not cultured in vitro prior to or after contact with the microcarriers, and wherein the majority of microcarriers within the plurality is porous.

3. The method of claim 1, wherein the cells are connective tissue progenitor cells.

4. The method of claim 1, wherein the cells are hematopoietic progenitor cells.

5. The method of claim 1, wherein the cells are endothelial progenitor cells.

6. The method of claim 1, wherein the sample is contacted with the plurality of microcarriers in a vessel.

7. The method of claim 1, wherein the sample and the plurality of microcarriers in solution are continuously stirred during the contacting step.

8. The method of claim 1, wherein the microcarriers are harvested from the solution using density based separation.

9. The method of claim 1, wherein the microcarriers are harvested from the solution using filtration.

10. The method of claim 1, wherein the sample and solution are in a volume ratio of about 1:

50.

11. The method of claim 1, wherein the sample is a freshly explanted sample.

12. The method of claim 1, wherein the sample is a tissue sample.

54. A method for transplanting cells into a subject comprising

isolating cells from a sample according to the method of claim 1, and

administering the cells into a subject in need thereof.

60. A kit comprising

a sterile vessel comprising an inlet/outlet port and a stirrer, and

instructions for isolating cells from a sample according to the method of claim 1, wherein the microcarriers are contacted with the sample in the vessel.

61. A kit comprising

a sterile vessel comprising an inlet/outlet port and a stirrer,

a first container comprising a plurality of sterile microcarriers,

a second container comprising one or more agents, and

a third container comprising a sterile solution, and optionally

instructions for isolating cells from a sample according to the method of claim 1.

66. A pharmaceutical composition comprising

cells prepared according to the method of claim 1 bound to one or more microcarriers, and

a pharmaceutically acceptable carrier.

2. A method for isolating cells from a sample comprising

contacting a sample with a plurality of microcarriers in solution under conditions and for a time sufficient for cells in the sample to bind to the microcarriers, and

harvesting from the solution microcarriers bound to cells,

wherein the sample is not cultured prior to or after contact with the microcarriers, and the majority of microcarriers in solution is not in contact with more than one other microcarrier.

13-53. -53. (canceled)

55-59. -59. (canceled)

62-65. -65. (canceled)

67-74. -74. (canceled)

75. A non-emulsion method for making a fibrin microcarrier composition, comprising

combining fibrinogen and thrombin to form a fibrin hydrogel,

dehydrating the fibrin hydrogel, and

grinding the dehydrated fibrin hydrogel to produce fibrin microcarriers.

98. A method for producing cell-bound fibrin microcarriers comprising

contacting a fibrin microcarrier made according to the method of claim 75 with a cell for a time and under conditions sufficient for the cell to bind to the fibrin microcarrier.

76-83. -83. (canceled)

84. A composition comprising

a plurality of fibrin microcarriers, at least 50% of which have an average diameter of 85-180 μ

m prior to hydration.

99. A method for producing cell-bound fibrin microcarriers comprising

contacting a population of fibrin microcarriers of claim 84 with cells for a time and under conditions sufficient for the cells to bind to the fibrin microcarriers.

85-97. -97. (canceled)

100-108. -108. (canceled)