Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Methods and compositions for inducing hematopoietic cell differentiation

Inactive Publication Date: 2018-03-15
FATE THERAPEUTICS
View PDF5 Cites 24 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patent describes a method for efficiently differentiation pluripotent stem cells into various types of blood cells such as T, B, NKT, and NK cells. This is done using a monolayer differentiation platform which combines high differentiation efficiency with large-scale expansion. The resulting blood cells have been shown to be functional and capable of long-term hematopoietic self-renewal, reconstitution, and engraftment in vivo. Overall, this patent provides a reliable and scalable method for generating therapeutically relevant amounts of blood cells from pluripotent stem cells.

Problems solved by technology

The methods provided herein for obtaining cells of the hematopoietic lineage are superior to EB-mediated pluripotent stem cell differentiation, because EB formation leads to modest to minimal cell expansion, does not allow monolayer culturing which is important for many applications requiring homogeneous expansion, and homogeneous differentiation of the cells in a population, and is laborious and low efficiency.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Methods and compositions for inducing hematopoietic cell differentiation
  • Methods and compositions for inducing hematopoietic cell differentiation
  • Methods and compositions for inducing hematopoietic cell differentiation

Examples

Experimental program
Comparison scheme
Effect test

example 1

hiPSC Generation and Maintenance

[0314]Somatic cells including fibroblast and blood cells were induced to reprogram towards a pluripotent state using various factor combinations including OCT4 / SOX2 / LargeT, OCT4 / SOX2 or OCT4 / SOX2 / NANOG / LargeT in the presence of reprogramming medium containing ROCK, MEK, GSK3 pathway and TGFβ receptor inhibitors (Valamehr et al. Sci Rep. 2012; 2: 213). Fourteen days after induction, reprogramming populations were switched to maintenance medium containing ROCK, GSK3, and MEK pathway inhibitors, basic fibroblast growth factor (bFGF), and leukemia inhibitory factor (LIF) (Valamehr et al. Stem Cell Reports 2014, 2(3): 366-381). Cells were kept indefinitely in the maintenance medium.

[0315]Approximately three weeks after induction, the reprogramming populations were sorted into individual wells of a 96-well plate. Selected clones were characterized and fully reprogrammed clones representative of naïve hiPSCs were selected for differentiation studies (Valameh...

example 2

Hematopoietic Differentiation Using iHSC Culture Platform

[0316]To initiate differentiation towards hematopoietic cell lineage, naïve hiPSCs were seeded as a monolayer in the maintenance medium and allowed to expand until approximately 25% confluency was reached. At this point, hematopoietic differentiation was initiated by switching the culture medium to iHSC-A (see FIG. 1). As illustrated in FIG. 1, the culture was subsequently switched to iHSC-B, 48hrs post the initiation of differentiation, followed by a switch to iHSC-C on day 4-5 post initiation of differentiation. The attached culture was maintained adherent and not perturbed during the medium changes.

[0317]Early during the differentiation process, directed differentiation was monitored by lineage markers, CD57, NESTIN, SOX17 and BRACHYURY. FIG. 3 illustrates a directed differentiation shift towards the mesoderm lineage and away from ectoderm lineage. Through the subsequent differentiation stages, the hiPSC morphology was give...

example 3

In Vitro Characterization and Testing Following Differentiation Using iHSC Culture Platform

[0318]To determine the expandability and maintenance of the differentiated definitive hematopoietic stem cells, the CD34 sorted or enriched population was transferred into suspended culture supplemented with Stemspan hematopoietic stem cell culture medium (StemCell Technologies, Vancouver, Canada), 1× CC110 supplement (StemCell Technologies), 10 ng / mL bFGF and 5 μM Thiazovivin or 10 μM 27632 ROCK inhibitors (for the first few days in culture to improve survival). The culture was fed with fresh medium every other day and pipetted to break up the aggregates resulting from dividing CD34 positive sorted cells. After several weeks in culture, the scaled suspended culture was assessed and measured through surface marker expression and viable cell number. As demonstrated in FIG. 8, the CD34 sorted population was maintained for 22 days in culture with minimal loss of the CD34 population. To improve vi...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides culture platforms, cell media, and methods of differentiating pluriptent cells into hematopoietic cells. The invention further provides pluripotent stem cell-derived hematopoietic cells generated using the culture platforms and methods disclosed herein, which enable feed-free, monolayer culturing and in the absence of ER formation. Specifically, pluripotent stem cell-derived hematopoietic cell of this invention include, and not limited to, iHSC, definitive hemogenic endothelium, hematopoietic multipotent progenitors, T cell progenitors, NK cell progenitors, T cells, and NK cells.

Description

RELATED APPLICATION[0001]This application claims priority to U.S. Provisional Application Ser. No. 62 / 107,517, filed Jan. 26, 2015 and U.S. Provisional Patent Application No. 62 / 251,016, filed Nov. 4, 2015, the disclosures of which are hereby incorporated by reference in their entirety.FIELD OF INVENTION[0002]The invention relates generally to compositions and methods for manufacturing cells of all hematopoietic lineages from pluripotent stem cells. In particular, the invention relates to improved culture platforms for manufacturing cells of all hematopoietic lineages from pluripotent stem cells including human induced pluripotent stem cells.BACKGROUND[0003]Human induced pluripotent stem cell (hiPSC) technology represents a highly promising and potentially unlimited source of therapeutically viable hematopoietic cells for the treatment of numerous hematological and non-hematological malignancies including cancer. To advance the promise of hiPSC and genomically engineered hiPSC techn...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/0789C12N5/0783
CPCC12N5/0636C12N5/0647C12N2501/26C12N2501/2311C12N2501/2306C12N2501/125C12N2501/165C12N2501/14C12N2501/22C12N2501/2303C12N2501/145C12N2501/727C12N2506/02C12N2501/415C12N2501/115C12N2501/155C12N5/0646C12N2506/11C12N2506/45C12N2501/105C12N2500/38C12N2500/02C12N2501/2302C12N2501/2307C12N2501/2315C12N2533/52C12N2506/1369C12N2501/998C12N2501/999C12N2501/42C12N2533/90A61P37/02C12N5/0018C12N5/0607C12N5/0662C12N5/0696C12N2501/10C12N2501/23C12N2501/40C12N2501/235
Inventor VALAMEHR, BAHRAMCLARKE, RAEDUN
Owner FATE THERAPEUTICS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com