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Compositions and methods for inducing myeloid suppressive cells and use thereof

A myelosuppressive, compositional technology applied in the field of adoptive immune cell therapy, which can solve the problems of inconsistent cell content, uneven exposure, hindering the scalability and reproducibility of the manufacturing process, etc.

Pending Publication Date: 2020-03-24
FATE THERAPEUTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

EB formation has been shown to promote pluripotent stem cell differentiation, however the requirement to form aggregates and subsequent EBs is labor-intensive with minimal increase in cell number during the process and the cell content in three-dimensional EB aggregates is inconsistent and variable. Uniform exposure to media factors, which yields heterogeneously derived cells at variable stages of differentiation and greatly hinders scalability and reproducibility requiring efficient, consistent, and streamlined manufacturing processes

Method used

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  • Compositions and methods for inducing myeloid suppressive cells and use thereof
  • Compositions and methods for inducing myeloid suppressive cells and use thereof
  • Compositions and methods for inducing myeloid suppressive cells and use thereof

Examples

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example

[0172] The following examples are provided by way of illustration and not by way of limitation.

example 1

[0173] Example 1 - Production of myeloid-derived suppressor cells from induced pluripotent stem cells

[0174]To initiate differentiation towards the hematopoietic lineage, hiPSCs were plated on day 0 (D0) as a monolayer in maintenance medium containing small molecules, including ROCK inhibitors, MEK inhibitors, and GSK3 inhibitors, and all The hiPSCs adhered and expanded for about 24 hours. EBs are not formed during this process. On D1, the maintenance medium was removed and replaced with basal medium (e.g., containing StemPro 34, glutamine, non-essential amino acids (NEAA), ascorbic acid, and monothioglycerol (MTG) without combining with small molecules. )) replacement. Around D2, hematopoietic differentiation was initiated by switching the medium to iCD34-A (eg, including basal medium and BMP4). At D3, the medium was supplemented with the growth factor bFGF and then switched to iCD34-B medium (eg, including basal medium, BMP4, bFGF, and GSK3 inhibitor) for differentiatio...

example 2

[0183] Example 2—iMDSCs potently inhibit T cell proliferation and effector function independently of HLA matching

[0184] To assess the function of iMDSCs, 5 × 10 4Ficoll-isolated PBMCs previously frozen and stored were left overnight, labeled with Cell Trace Violet (Invitrogen, Carlsbad, CA) and treated with anti-CD3 / 28 beads (ThermoFisher) activation. iMDSCs were co-cultured with activated PBMCs at a ratio of 1:1, 1:2 or 1:4 in complete RPMI medium in 96-well U-bottom plates. Five days later, co-cultures were harvested, and T cell expansion was quantified by flow cytometry. PBMC / iMDSC co-cultures were washed, stained with Live / Dead immobilized near-infrared viability dye (eBioscience) to exclude dead cells, and FC blocked (BD Biosciences, San Diego, CA) on ice. Biosciences, San Diego, Ca)) for about 30 minutes and surface stained on ice with fluorescent conjugated antibodies (BD Biosciences Format and Biolegend, San Diego, CA) for about 30 minutes to CD3(UCHT1), CD3(UCH...

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Abstract

Compositions and methods for manufacturing induced immune regulatory cells comprising induced myeloid suppressive cells including MDSCs (myeloid-derived suppressor cells), dendritic cells, macrophages, and subpopulations thereof are provided. Also provided are methods and compositions for further modifying and modulating the induced immune regulatory cells to achieve enhanced therapeutic potentialin treating autoimmune disorders, hematological malignancies, solid tumors, viral infections, neurodegenerative disorders, inflammatory conditions, or GvHD.

Description

[0001] related application [0002] This application claims priority to US Provisional Application Serial No. 62 / 519,123, filed June 13, 2017, the disclosure of which is hereby incorporated by reference in its entirety. technical field [0003] The present disclosure relates broadly to the field of adoptive immune cell therapy. More specifically, the present invention relates to an improved culture platform for the production of derived regulatory immune cells of the myeloid lineage suitable for adoptive cell therapy from pluripotent stem cells, including human induced pluripotent stem cells. Background technique [0004] Adoptive immunotherapy involves the administration of immune cells to a patient with a tumor, cancer, immune disorder or infection whereby the administered immune cells provide a therapeutic benefit to the patient. In general, immune cells suitable for immunotherapy include, but are not limited to, B cells, T cells, natural killer (NK) cells, NKT (natural...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078A61K35/15
CPCA61K39/461A61K39/4621A61K39/46434A61K35/44C12N5/0634C12N2500/02C12N2501/115C12N2501/125C12N2501/155C12N2501/165C12N2501/22C12N2501/2303C12N2501/2306C12N2501/2311C12N2501/26C12N2501/727C12N2501/999C12N2502/11C12N2506/45C12N2510/00C12N2533/50A61K2239/38A61K2239/31
Inventor P·A·帕龙R·S·塔科B·瓦拉梅尔D·舒梅克M·霍斯金L·格雷塔兹
Owner FATE THERAPEUTICS
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