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Method for differentiation of human pluripotent stem cell lines in suspension culture

A technology of pluripotent stem cells and cells, applied in the direction of artificially induced pluripotent cells, biochemical equipment and methods, animal cells, etc., can solve unproven problems such as scaling up

Pending Publication Date: 2020-12-04
AGENCY FOR SCI TECH & RES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although many methods for differentiating human induced pluripotent stem cells into erythroid cells have been described, these methods have not proven to be scalable

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  • Method for differentiation of human pluripotent stem cell lines in suspension culture
  • Method for differentiation of human pluripotent stem cell lines in suspension culture
  • Method for differentiation of human pluripotent stem cell lines in suspension culture

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Embodiment Construction

[0054] O-Negative Rhesus factor D negative (O-neg) blood is a common donor blood type and is considered a limited and valuable source of red blood cells (RBC) for emergency transfusion applications. Anticipated future supply shortages due to an aging population and the risk of emerging viruses and pathogens have driven initiatives to develop alternative and readily available universal sources of donor blood. The potential of using O-neghiPSCs as a starting material to generate universal donor erythrocytes has been a long-standing idea and has recently been demonstrated. The unlimited proliferation potential of hiPSCs and their potential to differentiate into hematopoietic lineage 4 have made these cells an unlimited source of cells for the generation of universal erythrocytes. Presumably, as few as 10 hiPSC clones with rare blood types are sufficient to cover 99% of the population requiring repeated transfusions. With the generation of high-quality GMP-grade hiPSCs and the co...

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Abstract

Described herein is a method of differentiation of pluripotent stem cells into hematopoietic precursor cells, hematopoietic progenitor cells, erythroblast cells, and enucleated erythroblast cells, wherein the method is carried out under suspension agitation, and a GSK-3-inhibitor or a Wnt pathway activator is added during a stage of mesoderm induction. In the preferred embodiment, the pluripotentstem cells are expanded in agitated microcarrier culture and the GSK-3-inhibitior is CHIR99021. Also disclosed herein are cell culture media for use in the methods disclosed herein, as well as kits for performing the same.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of priority to SG Provisional Application No. 10201800488W filed January 18, 2018, the entire contents of which are incorporated herein by reference for all purposes. technical field [0003] The present invention relates generally to the field of molecular biology. In particular, the invention relates to methods of cell differentiation. Background technique [0004] Erythroid differentiation of human induced pluripotent stem cells (hiPSCs) has been proposed as a means to generate an unlimited supply of red blood cells (RBCs). Therefore, there is a need to develop scalable suspension culture differentiation methods. Erythroid differentiation of human pluripotent stem cells (hPSCs) expanded in agitated microcarrier (MC) suspension cultures was previously shown using a bone morphogenetic protein-4 (BMP4)-based differentiation protocol in which mesoderm induction and erythroblast expa...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/07
CPCC12N2506/45C12N5/0696C12N2506/11C12N2510/00C12N2501/603C12N2501/602C12N2501/604C12N2501/606C12N2501/155C12N2506/02C12N5/0647C12N2501/115C12N2501/165C12N2501/16C12N2501/415C12N2501/392C12N2501/39C12N2501/2303C12N2501/14C12N2501/26C12N2501/145C12N2501/105C12N2501/998C12N2501/999C12N5/0641C12N2501/10C12N2501/15C12N2501/20C12N2501/727C12N2506/00C12N2527/00
Inventor J·斯瓦林加姆胡家荣S·鲁文尼
Owner AGENCY FOR SCI TECH & RES
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