Treatment of peripheral vascular disease using postpartum-derived cells

A peripheral vascular disease, cell-based technology for cell-based or regenerative therapy that addresses issues such as insufficient cord blood volume or cell numbers, cell population heterogeneity, and lack of characterization

Inactive Publication Date: 2009-05-20
ETHICON INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Limiting factors for obtaining stem cells from these methods have been the insufficient volume or number of cells obtained from cord blood, and the heterogeneity or lack of characterization of cell populations obtained from these sources

Method used

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  • Treatment of peripheral vascular disease using postpartum-derived cells
  • Treatment of peripheral vascular disease using postpartum-derived cells
  • Treatment of peripheral vascular disease using postpartum-derived cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0133] Umbilical cord cell culture and isolation Umbilical cord-derived cells (UDCs) were prepared according to US Patent Publication No. 2005 / 0058631 or 2005 / 0054098. Cells were cultured to passage 10 or 11 (approximately 20-25 population doublings) and then stored frozen.

[0134] Ischemia model treatment group:

[0135] Sample preparation for injection Cells were thawed immediately before injection (groups 3-5) or cultured for 24-30 hours (group 6). Cells were counted and viability was determined by trypan blue staining and counting with a hemocytometer. The entire dose of cells or plasmids (100 μg) was resuspended in 100 μl PBS and loaded into a 300 μl tuberculin syringe with a 27 gauge needle for injection into mice.

[0136] Surgery On day 0, acute hindlimb ischemia was induced in athymic nude mice by surgical unilateral ligation and resection of the left iliofemoral artery. Mice were divided into 6 groups of at least n=8 for treatment with UDCs or controls. For grou...

Embodiment 2

Endothelial network formation assay

[0144] Angiogenesis, or the formation of new blood vessels, is required for the growth of new tissue. Induction of angiogenesis is an important therapeutic goal in many pathological conditions. To identify the potential angiogenic activity of postpartum-derived cells in an in vitro assay, an established method was followed: Endothelial cells were seeded on plates coated with a biological cell culture substrate, MATRIGEL (BD Discovery Labware, Bedford, MA), a basement membrane extract (Nicosia and Ottinetti (1990) In Vitro Cell Dev. Biol. 26(2):119-28). Treatment of endothelial cells on MATRIGEL (BD Discovery Labware, Bedford, MA) with angiogenic factors stimulates the cells to form a network resembling capillaries. This is a general in vitro assay for testing stimulators and inhibitors of angiogenesis (Ito et al. (1996) Int. J. Cancer 67(1):148-52). The experiments utilized this co-culture system in which postpartum-derived cells were ...

Embodiment 3

Effects of hUTCs on proliferation and migration of endothelial cells in vitro

[0158] Studies were performed to determine the effect of human umbilical cord tissue-derived cells (hUTCs) on the proliferation and migration of endothelial cells in vitro. These effects were investigated by co-culturing hUTCs with endothelial cells and by incubating cultures of human umbilical vein endothelial cells (HUVECs) with hUTC lysates. The results presented here show that hUTCs induce increased proliferation and migration of endothelial cells. Furthermore, the data also suggest that these effects are mediated in part by fibroblast growth factor (FGF) and hepatocyte growth factor (HGF).

Materials and methods

cell culture

[0159] Refrigerated human umbilical cord tissue-derived cells (hUTCs) of lot number 120304 were thawed between passages 8-9, inoculated into gelatin-coated bottles, and grown in Hayflick growth medium (DMEM-low glucose [Gibco, catalog number 11885-084] , 15% v / v ...

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Abstract

Compositions and methods of using cells derived from postpartum tissue such as the umbilical cord and placenta, to stimulate and support angiogenesis, to improve blood flow, to regenerate, repair, and improve skeletal muscle damaged by a peripheral ischemic event, and to protect skeletal muscle from ischemic damage in peripheral vascular disease patients are disclosed.

Description

[0001] Cross References to Related Applications [0001] This application claims the benefit of US Provisional Patent Application Serial No. 60 / 754,366, filed December 28, 2005, the contents of which are hereby incorporated by reference in their entirety. field of invention [0002] The present invention relates to the field of cell-based therapy or regenerative therapy for patients with peripheral vascular disease, especially peripheral ischemia. In particular, the present invention provides postpartum tissue-derived cells capable of stimulating and supporting angiogenesis, improving blood flow, regenerating, repairing, and improving skeletal muscle damaged by peripheral ischemic events, and protecting skeletal muscle from deficient blood damage. Background of the invention [0003] Throughout this specification, reference is made to various publications, including patents, published applications, technical articles, and scholarly articles. Each of these cited publication...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P9/10C12N5/06A61K35/48A61K35/50C12N5/073
CPCA61L27/3886A61L27/3895C12N2510/00A61L27/3804A61L27/3839A61K35/50C12N5/0605A61K35/48A61P9/00A61P9/10A61P21/00A61P43/00
Inventor A·J·金
Owner ETHICON INC
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