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Repair and regeneration of renal tissue using human umbilical cord tissue-derived cells

A technology of umbilical cord tissue, human umbilical cord, used in the field of cell-based therapy

Inactive Publication Date: 2011-04-27
ETHICON INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, no therapeutic intervention attempts to halt or even reverse the progression of kidney disease

Method used

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  • Repair and regeneration of renal tissue using human umbilical cord tissue-derived cells
  • Repair and regeneration of renal tissue using human umbilical cord tissue-derived cells
  • Repair and regeneration of renal tissue using human umbilical cord tissue-derived cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] Isolation of Umbilical Cord Tissue-Derived Cells

[0103]Umbilical cords were obtained from National Disease Research Interchange (NDRI, Philadelphia, PA). These tissues were obtained after normal delivery. The cell isolation protocol was performed under sterile conditions in a laminar flow clean bench. To remove blood and debris, the umbilical cord was treated with antifungals and antibiotics (100 units / ml penicillin, 100 μg / ml streptomycin, 0.25 μg / ml amphotericin) in phosphate-buffered saline (PBS; Invitrogen, Carlsbad, CA). Wash in the presence of prime B). These tissues were then placed in a 150cm 2 Mechanical dissociation was performed in tissue culture dishes in the presence of 50 ml medium (DMEM-low glucose or DMEM-high glucose, Invitrogen) until the tissue was finely minced. Transfer the minced tissue to 50 mL conical tubes (approximately 5 grams of tissue per tube). The tissue is then digested in DMEM-low glucose medium or DMEM-high glucose medium, each...

Embodiment 2

[0109] Evaluation of human postpartum-derived cell surface markers by flow cytometry

[0110] Umbilical cord tissue was characterized using flow cytometry to provide a map for identification of cells obtained therefrom.

[0111] Cells were cultured in growth medium (Gibco Carlsbad, CA) containing penicillin / streptomycin. Cells were cultured in plasma-treated T75, T150 and T225 tissue culture flasks (Corning Corning, NY) until confluent. The growth surface of the flask was covered with gelatin by incubating 2% (w / v) gelatin (Sigma, St. Louis, MO) for 20 minutes at room temperature.

[0112] Adherent cells in culture flasks were washed in PBS and detached with trypsin / EDTA. Cells were collected, centrifuged and separated at 1×10 7 The concentration of cells per milliliter was resuspended in 3% (v / v) FBS in PBS. Antibodies to cell surface markers of interest (see below) were added to one hundred microliters of the cell suspension according to the manufacturer's instructions...

Embodiment 3

[0137] Immunohistochemical identification of cellular phenotypes

[0138] Human umbilical cord tissue was collected and immersion fixed in 4% (w / v) paraformaldehyde at 4°C overnight. Immunohistochemistry was performed using antibodies directed against the following epitopes: vimentin (1:500; Sigma, St. Louis, MO), desmin (1:150, raised against rabbit; Sigma; or 1:300, against Increased in mice; Chemicon, Temecula, CA), α-smooth muscle actin (SMA; 1:400; Sigma), cytokeratin 18 (CK18; 1:400; Sigma), von Willebrand factor (vWF; 1: 200; Sigma) and CD34 (human CD34 class III, 1:100, DAKOCytomation, Carpinteria, CA). In addition, the following markers were tested: anti-human GROα-PE (1:100; Becton Dickinson, Franklin Lakes, NJ), anti-human GCP-2 (1:100; Santa Cruz Biotech, Santa Cruz, CA), anti-human oxidative LDL receptor 1 (ox-LDL R1; 1:100; Santa Cruz Biotech) and anti-human NOGO-A (1:100; Santa Cruz Biotech). Fixed samples were trimmed with a scalpel and placed in OCT embed...

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Abstract

Methods for treating a patient having a disease or damage to at least one kidney are provided. The methods comprise administering cells obtained from human umbilical cord tissue, or administering pharmaceutical compositions comprising such cells or prepared from such cells. When administered, the cells promote and support the repair and regeneration of the diseased or damaged kidney tissue in the patient. Pharmaceutical compositions for use in the inventive methods, as well as kits for practicing the methods are also provided.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to US Provisional Patent Application No. 60 / 977775, filed October 5, 2007, the entire contents of which are hereby incorporated by reference. technical field [0003] The present invention relates generally to the field of cell-based therapies. More specifically, the present invention relates to the use of umbilical cord tissue-derived cells in the repair and regeneration of diseased or damaged kidneys. technical background [0004] Various publications are cited throughout this specification, including patents, published applications, technical articles, and scholarly articles. All of these cited publications are hereby incorporated by reference in their entirety for various purposes. [0005] Kidney disease is a serious, medically unmet condition with an annual cost burden of more than $27 billion in the United States. Currently, more than 40 million Americans are at risk for or li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00C12N5/08A61K35/12A61K35/44C12N5/073
CPCC12N5/0605A61K35/44A61K35/12A61P13/12
Inventor D·科尔特A·戈谢夫卡
Owner ETHICON INC
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