Regeneration of a functional pulmonary vascular bed

A pro-angiogenic, vascular technology for the regeneration of functional pulmonary vascular beds that can address issues that limit long-term outcomes

Active Publication Date: 2018-10-23
THE GENERAL HOSPITAL CORP
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the number of donors is not meeting the growing demand, while t...

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  • Regeneration of a functional pulmonary vascular bed
  • Regeneration of a functional pulmonary vascular bed
  • Regeneration of a functional pulmonary vascular bed

Examples

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example

[0128] The following specific examples further illustrate the present invention.

example 1

[0130] Study of the Vascular Bed of the Acellular Pulmonary

[0131] The purpose of this example was to characterize the perfusion properties of the acellular vascular bed in a whole lung scaffold using an in vitro microsphere perfusion assay. By quantifying microspheres harvested from the PV, trachea, and lung periphery, we demonstrate both the continuity and integrity of the vascular basement membrane after decellularization and thereby confirm compartment-specific cell delivery in acellular lung scaffolds possibility. Microsphere quantification further implicates fluid-driven hydrostatic pressure loss rather than particle leakage as the primary cause of low passage efficiency. By applying these findings to recellularization, efficient and uniform endothelial coverage is achieved through combined arterial and venous cell delivery.

[0132] method

[0133] Cadaveric lungs were removed from male Sprague-Dawley rats (250-300 g, Charles River Laboratories) following systemic ...

example 2

[0143] Improved endothelial delivery

[0144] The purpose of this example is the reconstitution of viable endothelial cells on cell-free rat lung scaffolds and confirmation of the integrity of the vascular basement membrane after decellularization.

[0145] method

[0146] Cell seeding into decellularized rat lungs was performed in bioreactors similar to those described herein, allowing cell delivery and perfusion from PA and PV. The trachea was cannulated and opened to the interior of the bioreactor through a port located approximately 5 cm above the PA level. Decellularized lung scaffolds were filled by perfusing with 100 ml of Hank's balanced salt solution containing human fibronectin (2.5 μg / ml) at 1 ml / min for 1 hour, washed with Hank's balanced salt solution for 1 hour, and Equilibrate in the respective media for at least 1 h before cell seeding. For endothelial delivery by PA, 40 million HUVECs were resuspended in a single inoculation chamber with 100 ml EGM-2 and in...

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Abstract

A method for vascular regeneration comprises delivering endothelial cells to a lung scaffold, delivering perivascular cells to the lung scaffold, and providing a multiphase culture program to the scaffold. The multiphase culture program comprises a first phase including delivering an angiogenic medium, e.g., having 40-100 ng/ml of pro-angiogenic factors, and a second phase including delivering a stabilization medium, e.g., having 0.5-2% of serum and 1-20 ng/ml of angiogenic factors.

Description

technical field [0001] The present disclosure relates to the regeneration of a functional pulmonary vascular bed. Background technique [0002] Lung transplantation is an effective treatment option for patients with end-stage lung disease. However, donor numbers are not meeting the growing demand, and the effects of chronic immunosuppression and rejection limit long-term outcomes. Typical wait times for lung transplants can be two years or more, resulting in a 30 percent mortality rate for those on the waiting list. Contents of the invention [0003] The present disclosure is based at least in part on the development of devices (e.g., bioreactors) and methods capable of regenerating functional pulmonary vessels by delivering cells and repopulating the vascular compartment of acellular lung scaffolds with endothelium and perivascular cells bed and mature the pulmonary vascular bed using a multi-stage culture procedure. Implementations can include one or more of the follo...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12M3/00
CPCC12N5/069C12N5/0688C12M21/08C12M25/14C12M41/48C12M29/04C12M29/10C12M41/40C12M33/04C12N2506/45C12N2501/135C12N2501/155C12N2501/415C12N2501/115C12N2533/92C12N2501/11C12N2501/01C12N2501/15C12N2500/25C12N2501/165C12N2500/02C12N2501/17C12N2501/39C12N2502/28C12N2502/1388C12N2502/27C12N5/0697A61P11/00C07K14/475
Inventor 任息H·C·奥特
Owner THE GENERAL HOSPITAL CORP
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