Perivascular mesenchymal precursor cells

• US 7,947,266 B2
• Filed: 03/29/2004
• Issued: 05/24/2011
• Est. Priority Date: 03/28/2003
• Status: Active Grant

• Current Assignee: Mesoblast, Inc. (Mesoblast Ltd.)
• Original Assignee: Angioblast Systems, Inc.
• Inventors: Shi, Songtao, Gronthos, Stan, Zannettino, Andrew
• Primary Examiner(s): Belyavskyi; Michail A
• Application Number:
  US10/551,162
• Publication Number:
  US 20070065938A1
• Time in Patent Office: 2,612 Days
• Field of Search: None
• US Class Current: 424/93.1
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 35/44 Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 9/0014 Skin, i.e. galenical aspects of topical compositions non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2500/42 Organic phosphate, e.g. beta glycerophosphate
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2506/1392 from mesenchymal stem cells from other natural sources
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0667 Adipose-derived stem cells [ADSC]; Adipose stromal stem cells
• CPC Class Codes: C12N 5/0668 Mesenchymal stem cells from other natural sources
• CPC Class Codes: C12N 5/0691 Vascular smooth muscle cells; 3D culture thereof, e.g. models of blood vessels

Abstract

Mesenchymal precursors cells have been isolated from perivascular niches from a range of tissues utilizing a perivascular marker. A new mesenchymal precursor cell phenotype is described characterized by the presence of the perivascular marker 3G5, and preferably also alpha smooth muscle actin together with early developmental markers such as MUC 18, VCAM-1 and STRO-1^bri. The perivascular mesenchymal precursor cell is multipotential and is shown to form, vascular tissue, as well as bone marrow dentin and pulp. A method of enriching using cell sorting based on these markers is also described.

Claims

1. A population of cells enriched for 3G5 cells, wherein such 3G5 cells are mesenchymal precursor cells which comprise mesenchymal precursor cells capable of giving rise to colony forming unit-fibroblast (CFU-F), and wherein at least 30% of the total cells of the population are positive for the marker 3G5.

2. The enriched population of claim 1 wherein the MPCs are enriched from a perivascular niche within a non-haemopoietic vascularised tissue.

3. The enriched population of claim 1 wherein the MPCs are enriched from a tissue of the group consisting of skin, liver, kidney, heart, adipose tissue, teeth, dental pulp, retina, brain, hair follicles, intestine, lung, spleen, lymph node, thymus, pancreas, bone, ligament, bone marrow, tendon and skeletal muscle.

4. The enriched population of claim 1 wherein the MPCs are also positive for one or more of the perivascular cell markers MUC18/CD146 and alpha-smooth muscle actin.

5. The enriched population of claim 1 wherein the enriched population comprises at least 0.1% STRO-1^bri MPCs.

6. The enriched population of claim 1 wherein the enriched population comprises at least 1% STRO-1^bri MPCs.

7. The enriched population of claim 1 wherein the MPCs are positive for the markers STRO-1^bri, MUC18/CD146, and alpha-smooth muscle actin.

8. The enriched population of claim 1 wherein the MPCs are positive for one or more markers selected from the group consisting of THY-1, VCAM-1, ICAM-1, PECAM-1, CD49a/CD49b/CD29, CD49c/CD29, CD49d/CD29, CD29, CD61, integrin beta 5, 6-19, thrombomodulin, CD10, CD13, SCF, STRO-1^bri, PDGF-R, EGF-R, IGF1-R, NGF-R, FGF-R and Leptin-R (STRO-2).

9. The enriched population of claim 1 wherein the MPCs are negative for the haemopoietic markers CD45, CD34, and glycophorin A.

10. The enriched population of claim 1 wherein the MPCs have the capacity to be induced to differentiate to form progeny cells comprising one or more of at least osteoblast, odontoblast, dentin-producing, chondrocyte, tendon, ligament, cartilage, adipocyte, fibroblast, marrow stroma, osteoclast- and hematopoietic-supportive stroma, cardiac muscle, smooth muscle, skeletal muscle, pericyte, vascular, epithelial, glial, neuronal, astrocyte or oligodendrocyte cell type.

11. An enriched population of claim 1 comprising at least 0.1% MPCs capable of forming a clonogenic colony.

12. An enriched population of claim 1 comprising at least 1% MPCs capable of forming a clonogenic colony.