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Amnion-derived cell compositions, methods of making and uses thereof

a technology of amnion-derived cells and compositions, applied in the field of amnion-derived cell compositions, can solve the problems of incisional hernia, failure to mechanically fix, and failure to successfully heal acute wounds,

Inactive Publication Date: 2006-10-05
STEMNION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0024] Although heterogeneous populations of placental-derived stem cells have been previously characterized using established embryonic stem cell surface protein markers such as c-kit, SSEA-3, and SSEA-4, a set of protein markers useful for characterizing and isolating a preferred substantially purified population of cells is required. This substantially purified population of cells, termed amnion-derived cells, could then be fully discriminated

Problems solved by technology

Subsequent acute wound healing failure at any anatomic site results in increased morbidity and mortality.
There are no other models of acute wound healing suggesting that a successfully healed acute wound goes on to breakdown and mechanically fail at a later date.
One model of laparotomy wound failure that was developed resulted in incisional hernias.
Diabetes—Traditional insulin therapy prolongs the life of a patient with Type I diabetes but does not prevent the long-term systemic complications that arise as the disease progresses.
Even the best injection / infusion regime to monitor and control systemic glucose levels within an acceptable range inevitably leads to a deterioration of tissue microvascularization resulting in the plethora of health-related complications associated with the disease.
These complications can be attributed to the inability of injectable or orally administered insulin to completely substitute for the insulin secretion from a normal complement of pancreatic islets.
The scarcity of donor tissue reserves these alternative therapies for select patients that are unable to stabilize their blood glucose adequately using traditional insulin injection / infusion regimes.

Method used

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  • Amnion-derived cell compositions, methods of making and uses thereof
  • Amnion-derived cell compositions, methods of making and uses thereof

Examples

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example 1

Preparation of Amnion-Derived Cell Compositions

[0271] Recovery of amnion-derived cells—Amnion-derived cells were dissociated from starting amniotic membrane using the dissociation agents PXXIII, and trypsin. The average weight range of an amnion was 18-27 g. The number of cells recovered per g of amnion was about 10-15×106 for dissociation with PXXIII and 5-8×106 for dissociation with trypsin.

[0272] Culture conditions—The primary amnion-derived cells were cultured for 5 passages in the following media: Stemline II+10% FBS, Stemline II+10% plasbumin (pb), Ultraculture+10% plasbumin (pb), and DMEM+10% FBS. Each culture condition was tested using 15 million cells / g amnion, 10 million cells / g amnion, and 5 million cells / g amnion, depending on the enzyme used for recovery of the primary cells. For instance, using PXXIII, 15 million cells / g amnion were obtained, while using trypsin, 10 million cells / g amnion were obtained, while other enzymes resulted in even lesser recovery (5 million ...

example 2

Scale-Up of Amnion-Derived Cells on Microcarrier Beads in Spinner Flasks

[0277] Methods—One of the most common and oldest techniques for maintaining cells in suspension culture is by the use of spinner flasks. The cells can be either attached to microcarrier beads (adherent) or growing completely without any surface attachment (non-adherent). In either case, these flasks consist of a sterile vessel that contains a magnetic stirring mechanism that permits continuous stirring of the medium and cells under sterile conditions. This continuous stirring facilitates the diffusion of nutrients, promotes oxygenation of the medium, and eliminates concentration gradients. The vessels are stirred in a temperature-controlled, CO2 incubator.

[0278] Amnion-derived cells are an epithelial cell type that are anchorage-dependent which may interfere with or prevent their adaptation to a pure suspension system. Although amnion-derived cells may survive in suspension culture, the proliferation of these ...

example 3

Scale-Up of Amnion-Derived Cell Compositions in Suspension

[0281] Amnion-derived cells were cultured in ultra-low adherence tissue culture 6-well plates (Corning) in various mammalian cell culture media. These culture media were selected on the basis of their ability to promote proliferation of other mammalian cell types in suspension culture (i.e. 293S, Ultraculture, Opti-MEM). Additives to the culture medium in these experiments include a proprietary source of protein, and EGF (10-20 ng / ml) which preliminary experiments show is required for proliferation of amnion-derived cells. Amnion-derived cells were plated at a density of 1.3×106 cells / well, and the cultures were maintained at 37° C., in 5% CO2 in air. Culture medium was replaced every two days and cell number was assessed weekly. Preliminary experiments showed that amnion-derived cells sometimes form small floating clusters in suspension culture conditions. These clusters must be dispersed to ensure accurate cell counts and ...

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Abstract

The invention is directed to substantially purified amnion-derived cell populations, compositions comprising the substantially purified amnion-derived cell populations, and to methods of creating such substantially purified amnion-derived cell populations, as well as methods of use. The invention is further directed to antibodies, in particular, monoclonal antibodies, that bind to amnion-derived cells or, alternatively, to one or more amnion-derived cell surface protein markers. The invention is further directed to methods for producing the antibodies, methods for using the antibodies, and kits comprising the antibodies.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority under 35 USC § 19(e) to U.S. Provisional Application No. 60 / 666,949, filed Mar. 31, 2005, U.S. Provisional Application No. 60 / 699,257, filed Jul. 14, 2005, U.S. Provisional Application No. 60 / 742,067, filed Dec. 2, 2005, and under 35 USC §120 to U.S. Utility application Ser. No. 11 / 333,849, filed Jan. 18, 2006, the contents of which are incorporated herein by reference in their entirety.FIELD OF THE INVENTION [0002] The field of the invention is directed to amnion-derived cell populations, compositions comprising the amnion-derived cell populations, expanded amnion-derived cell populations, methods of creating such amnion-derived cell populations, as well as methods of use. The field is also directed to antibodies, in particular, monoclonal antibodies, that bind to amnion-derived cells or, alternatively, to one or more amnion-derived cell surface protein markers, methods for producing the antibodies, met...

Claims

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Application Information

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IPC IPC(8): A01N63/00C12N5/00A61K35/12C12N5/071C12N5/073
CPCA61K35/12C12N5/0605C12N5/0629A61Q19/00C12N2502/02A61K35/50A61K8/982C12N2501/11A61K2800/10
Inventor CLARKE, DIANA L.SMITH, CHARLOTTE A.BANAS, RICHARD A.MARSHALL, VIVIENNE S.
Owner STEMNION
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