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Implantable constructs and uses thereof

A construct, entity technology, applied in biology, bioengineering and medical devices, medicine, which can solve the problem of lack of dual goals of controlled delivery of therapeutic agents

Pending Publication Date: 2022-04-01
RICE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, technologies to achieve the dual goals of FBR and controlled delivery of therapeutic agents are lacking

Method used

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  • Implantable constructs and uses thereof
  • Implantable constructs and uses thereof
  • Implantable constructs and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Example 1: Cell Engineering and Cell Culture

[0108] Reagents. Cell culture media and related reagents, such as transfection media, were purchased through Fisher Scientific. Lipofectamine ) and selection medium (puromycin) were purchased from Invitrogen. VectorBuilder develops and subsequently purchases expression vectors and helper plasmids. CellTiter- Purchased from Promega. Know Live Stains were purchased from Fisher Scientific. Alginate compounds were purchased from PRONOVA. The packaging equipment used contained a syringe pump from Harvard Apparatus and a coaxial nozzle from Ramé-Hart. All animals were purchased from Jackson Labs. Alginate compounds were purchased from PRONOVA.

[0109] Cell culture. ID8 / MOSEC cells were purchased from EMD Millipore, Sigma. PAN02 or PANC02 mouse cells were obtained from The Division of Cancer Treatment and Diagnosis (DCTD) of the National Cancer Institute. Human ARPE-19 cells were obtained from ATCC. ARPE- 19 cell...

Embodiment 2

[0114] Example 2: Quantitative production by ELISA

[0115] Cells from each group were grown in T-150 flasks in DMEM medium with 10% FBS, 1% Penstrep and 0.5 μl / 500ml puromycin. When confluent, the medium was aspirated and 10ml TrypLE was added. Place the cells in 5% CO 2 Incubate for 3 min at 37 °C in a humidified atmosphere. After the cell layer was dispersed, 10 ml of complete medium was added to stop the reaction. Transfer the cell suspension to a 50 ml conical tube and centrifuge at 250G for 5 min. Aspirate the supernatant and resuspend the cells in 1 ml complete medium. Use Countess TM A hemocytometer counts the cell concentration. Calculate the volume required to achieve a concentration of 10,000 cells in 200 µl based on the concentration of viable cells in the sample. In the remaining cell suspension volume, 1 million cells were resuspended in 1 ml alginate to synthesize core-shell capsules. Place all groups in 5% CO 2 Incubate for 24 hours at 37 °C in a humid...

Embodiment 3

[0116] Example 3: T cell proliferation assay

[0117] T cells were isolated from C57BL6 mouse spleens (Jackson Labs) using the EasySep T cell isolation kit (StemCell Technologies) and related reagents and devices. Cells were plated in 100ul RPMI 1640 at 10,000 cells / well in 96-well plates. Cells were supplemented with 0.1, 1, 5 or 10 ng / mL of native IL2 produced by cells or 1 or 10 ng / mL of recombinant IL-2 (Miltenyi). Cell proliferation (and viability) was measured using the CellTiter-Glo Luminescent Cell Viability Assay (Promega) after incubation at 37°C for 24 or 72 hours.

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Abstract

The present disclosure relates to implantable constructs designed to deliver an antigenic therapeutic agent to a subject while avoiding the effects of a host immune response. In certain aspects, the constructs are designed to degrade over time or after a particular signal, thereby controlling the duration of delivery of the therapeutic agent to the subject.

Description

[0001] priority claim [0002] This application claims the benefit of or priority to U.S. Provisional Application Serial Nos. 62 / 884,397 and 63 / 012,693, filed August 8, 2019, and April 20, 2020, respectively, each of which is incorporated by reference in its entirety In this article. [0003] Statement Regarding Federally Funded Research [0004] This invention was made with government support under Grant No. R01DK120459 awarded by the National Institutes of Health. The government has certain rights in this invention. technical field [0005] The present disclosure relates to the fields of biology, medicine, bioengineering and medical devices. More specifically, it relates to the development and use of implantable constructs designed to deliver antigenic therapeutic agents to subjects and protect them from host-generated immune responses . In particular, the constructs are designed to degrade over time or following specific signals, thereby controlling the length of time ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/69A61K9/50A61K9/00C12N5/00
CPCA61K38/00A61K35/00A61K9/0024A61K9/5036C07K14/54C07K14/5418C07K14/5434C07K14/5428C07K14/5443C07K14/55C12N2510/00C12N5/0012C12N2533/74A61P35/00A61K38/19A61K38/2013A61K38/2046A61K45/06
Inventor 奥米德·维塞阿曼达·纳什玛丽亚·伊莎贝尔·贾维斯-罗科苏迪普·慕克吉迈克尔·大卫·德费特萨米拉·阿格拉-福托瓦特大卫·于·张
Owner RICE UNIV
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