Stem cells from urine and methods for using the same

a technology of stem cells and urine, applied in the field of stem cells from urine, can solve problems such as limiting the supply of viable cells

Inactive Publication Date: 2010-05-06
WAKE FOREST UNIV HEALTH SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0019]Also provided are methods of treating stress urinary incontinence (UI) or vesicoureteral reflux (VCR) in a subject in need thereof, comprising administering urine stem cells to said subject in a treatment effective amount. In some embodiments, administering is carried out by subcutaneous administration. In some embodiments, administering is carried out by endoscopic injection. In some embodiments, urine stem cells are tranfected with a nucleic acid encoding VEFG. In some embodiments, the methods further comprise administering endothelial cells. In some embodiments, the cells are provided in a pharmaceutically acceptable carrier, e.g., a collagen gel, a hydrogel, a temperature sensitive gel or a hyaluronic acid gel.

Problems solved by technology

However, a donor shortage limits the supply of viable cells to use for these applications.

Method used

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  • Stem cells from urine and methods for using the same
  • Stem cells from urine and methods for using the same
  • Stem cells from urine and methods for using the same

Examples

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example 1

Isolation and Characterization of Stem Cells from Urine

[0127]Fifty-eight human urine samples were collected from 22 male and one female donors (15 healthy individuals and 8 patients), ranging in age from 2 to 50 years. No bacterial contamination was found in any cultures. Urine samples were centrifuged at 1500 RPM for 5 minutes at 4° C. and washed two times with sterile PBS. Cells were plated at an average of 0.5 cell / well in multi-well plates with stem cell medium using a gradual dilution method. The stem cell medium contains ¾ DMEM, ¼ Ham's F12, 10% FBS, 0.4 mg / ml hydrocortisone, 10−10 M, Chron Toxin, 5 mg / ml, insulin, 1.2 mg / ml adenine, 2.5 mg / ml transferrin plus 0.136 mg / ml 3,39,5-triiodo-L-thyronine, 10 mg / ml EGF, and 1% penicillin-streptomycin (Zhang et al., In vitro Cell Dev. Biol.-Animal 37:419, 2001). Single cells were identified and allowed to grow to more than 50% confluence. Cells were subsequently subcultured, transferred to a 6-cm culture dish and expanded.

[0128]Primar...

example 2

Cell Contractility and Tight Junctions of Urine Stem Cells

[0132]Functional characteristics of urine-derived smooth muscle cells are ascertained by determining cell contractility with collagen lattices. Methods for the collagen lattice contraction assay are known in the art (Kropp, et al. (1999) J. Urol. 162:1779). Contractile response to agonists is performed in a similar manner except that agonists are added to the serum-free media immediately prior to lattice release. Agonists which are tested include the Ca-ionophore A23187 (1025 M) and KCl.

[0133]Urine-derived urothelial stem cells are analyzed for tight junctions. Urothelial barrier function is maintained by apical membrane plaques and intercellular tight junctions. Tight junction components within urine stem cells is investigated with conventional methods such as electric microscopy (Zhang, et al. (2003) Adv. Exp. Med. Biol. 539:907; Ludwikowski, et al. (1999) BJU hit. 84:507; Sugasi, et al. (2000) J. Urol. 164:951; Zhang, et a...

example 3

Urine Stem Cell Differentiation into Smooth Muscle Cells

[0134]To determine the effect of epithelial-stromal cell interaction or cell-cell interaction on urine stem cell differentiation into smooth muscle cells (Staack, et al. (2005) Differentiation 73:121; DiSandro, et al. (1998) J. Urol. 160:1040), two co-culture methods are used. The first is an indirect co-culture using transwell devices with a 4.5 μm size exclusion. Urine smooth muscle stem cells are plated on the upper chamber wells and bladder smooth muscle cells and / or urothelium from tissue biopsy are seeded on the bottom wells of a transwell unit (Luk, et al. (2005) J. Immunol. Methods 305:39; Gerstenfeld, et al. (2003) Connect Tissue Res. 44(suppl 1):85). The second method is a direct co-culture approach which is performed with a transwell insert of 0.4 μm pore size. The transwell is used as a basement membrane on which urine stem cells are cultured on the lower side, while normal human bladder urothelium and smooth muscle...

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Abstract

Provided herein are stem cells and methods for producing a culture of stem cells from urine. The stem cells may be differentiated into an osteogenic, chondrogenic, adipogenic, endothelial, neurogenic or myogenic lineage. Methods of use of the cells are provided, including methods of treating a subject in need of a cell based therapy.

Description

RELATED APPLICATIONS[0001]This application claims priority to under 35 U.S.C. §120, and is a continuation-in-part of, PCT application no. PCT / US2008 / 006438, filed May 20, 2008, which claims the benefit under 35U.S.C. §119(e) of U.S. Provisional Patent Application Ser. No. 60 / 939,247, filed May 21, 2007, and U.S. Provisional Patent Application Ser. No. 60 / 943,215, filed Jun. 11, 2007, the disclosure of each of which is incorporated herein by reference in its entirety.[0002]This application also claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Patent Application Ser. No. 61 / 120,224, filed Dec. 5, 2008, and U.S. Provisional Patent Application Ser. No. 61 / 172,444, filed Apr. 24, 2009, the disclosure of each of which is incorporated herein by reference in its entirety.FIELD OF THE INVENTION[0003]This invention relates generally to the isolation of stem cells from urine, cells isolated, differentiation thereof into multiple lineages, and methods of use of the same.BACKGROUND...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K35/12C12N5/00A61P13/02A61K35/22
CPCA61K35/22C12N5/0684C12N5/0668A61P13/02
Inventor ZHANG, YUANYUANATALA, ANTHONY
Owner WAKE FOREST UNIV HEALTH SCI INC
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