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Augmentation of organ function

一种器官、功能的技术，应用在细胞群增强器官功能领域，能够解决出现感染、削弱免疫系统等问题

Inactive Publication Date: 2005-05-11
CHILDRENS MEDICAL CENT CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Prolonged immunosuppression may weaken the immune system, which may lead to the threat of infection

Method used

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  • Augmentation of organ function
  • Augmentation of organ function

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] Example 1: Isolation of kidney cells

[0138] Small kidneys, such as those from one-week-old C7 black mice, were capsuled, dissected, and ground, and suspended in Dulbecco's Modified Eagles' Medium (DMEM; Sigma, St. Louis, MO), which cultured The base contained 15 mM Hepes at pH 7.4, and 0.5 μg / ml insulin, 1.0 mg / ml collagenase, and 0.5 mg / ml dispase, a neutral protease from Bacillus polymyxa (Boehringer Mannheim, Indianapolis, IN).

[0139] Large kidneys, such as porcine kidneys, are arterially perfused with Eagles minimal essential medium without calcium for 10 minutes at 37°C, within 3 hours of extraction. Then, supplemented with 1.5mM MgCl 2 and 1.5mM CaCl 2 The kidneys were perfused with 0.5 mg / ml collagenase (Type IV, Sigma, St. Louis, MO) in the same buffer as that used for the perfusion. The kidneys were then stripped, dissected, crushed, and suspended in Dulbecco's modified Eagles' medium (DMEM; Sigma, St. Louis, MO) containing 15 mM Hepes at pH 7.4, and 0.5...

Embodiment 2

[0141] Embodiment 2: the in vitro culture of kidney cell

[0142] I. Isolation of Rat Tail Collagen

[0143] Tendons were dissected from rat tails and stored in 0.12M acetic acid solution in deionized water in 50 ml test tubes. After 16 hours, overnight at 4°C.

[0144] Dialysis bags are pretreated to ensure uniform pore size and removal of heavy metals. Briefly, dialysis bags were soaked in a solution of 2% sodium bicarbonate and 0.05% EDTA and boiled for 10 minutes. Sodium bicarbonate and 0.05% EDTA were removed by rinsing multiple times with distilled water.

[0145] A 0.12M acetic acid solution including rat tendon was placed into a treated dialysis bag and dialyzed over a period of 2-3 days to remove the acetic acid. Change the dialysis solution every 3-4 hours.

[0146] (ii) Coating the tissue culture plate:

[0147] With about 30 μg / ml collagen (Vitrogen or rat tail collagen), about 10 μg / ml human fibronectin (Sigma, St.Louis, MO) and about 10 μg / ml bovine serum a...

Embodiment 3

[0151] Example 3: Isolation and culture of endothelial cells

[0152] Isolate endothelial cells from dissected veins. An intravenous heparin / papaverine solution (3 mg papaverine hydrochloride diluted in 25 ml Hanks' balanced salt solution (HBSS) containing 100 units of heparin (final concentration 4 μg / ml)) was used to improve endothelial cell preservation. The proximal filamentary loop is placed around the venous vessel and secured with a knot. A small phlebotomy is made close to the node and the tip of the venous cannula is inserted and held in place with a second node. A second small phlebotomy was performed beyond the proximal segment and treated with 20% fetal bovine serum, ECGF (100mg / ml), L-glutamine, heparin (Sigma, 17.5u / ml) and antibiotic-antimycotic Medium 199 / Heparin Solution Medium 199 (M-199) to gently flush the vein to remove blood and blood clots. With about 1ml collagenase solution (0.2% Worthington type I collagenase dissolved in 98ml M-199, 1ml FBS, 1ml ...

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Abstract

The present invention provides methods and compositions for augmenting organ functions using small-scale matrix implants generated by seeding tissue-specific or undifferentiated cells onto a matrix materials (e.g., a wafer, sponge, or hydrogel). The seeded matrix composition can then be implanted and will develop into an organ-supplementing structure in vivo. Continued growth and differentiation of the seeded cells on the implanted matrix results in the formation of a primitive vascular system in the tissue. The primitive vascular system can then develop into a mature vascular system, and can also support the growth and development of additional cultured cell populations. The seeded matrix system can be used to introduce a variety of different cells and tissues in vivo.

Description

[0001] References to related applications [0002] This application claims priority to US Provisional Application Serial No. 60 / 331500, filed November 16, 2001. This application is also a continuation-in-part of US Patent Application Serial No. 09 / 474525, filed December 29, 1999. The contents of the above two related applications are expressly incorporated herein by reference. Background of the invention [0003] The technical field of the invention is the enhancement of organ function by transplantation of cultured cell populations. Often, much of any organ function may have been lost before a patient develops complete organ failure. For example, up to 90-95% of kidney function may have been lost before kidney failure manifests. This confirms the enormous capacity for rapid self-renewal (Cuppage et al., (1969) Lab.Invest.21:449-459; Humes et al., (1989) J Clin.Invest.84:1757-1761; and Witzgall et al. (1994) J. Clin. Invest. 93:...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00A61K35/12A61L27/00A61L27/38C12N5/00C12N5/071
CPCA61F2/022A61K35/12A61L27/3641A61L27/3683A61L27/3839A61L27/3886A61L2430/26C12N5/0068C12N5/0685C12N5/0686C12N2501/11C12N2502/28C12N2503/04C12N2533/52C12N2533/54C12N2533/90A61P13/12A61L27/38A61K45/00C12N5/0602
Inventor A·阿塔拉
Owner CHILDRENS MEDICAL CENT CORP
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