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Populations of hematopoietic progenitors and methods of enriching stem cells therefor

Inactive Publication Date: 2014-09-17
UNIV HEALTH NETWORK +1
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Problems solved by technology

Although these methods yield a broad class of hematopoietic progenitor cells, transplantation of progeny from such cultures into immunocompromised mice generally results in low-level engraftment that is often restricted to the myeloid lineage (Lu et al., 2009; Tian et al., 2006; Wang et al., 2005)
These findings suggest that conditions used for hematopoietic differentiation do not support HSC development

Method used

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  • Populations of hematopoietic progenitors and methods of enriching stem cells therefor
  • Populations of hematopoietic progenitors and methods of enriching stem cells therefor
  • Populations of hematopoietic progenitors and methods of enriching stem cells therefor

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Embodiment Construction

[0058] Materials and methods

[0059] Maintenance and differentiation of human ES and iPS cells

[0060] The hESC line H1 (Thomson et al., 1998) and a reprogrammed human iPS cell line (MSC-iPS1; (Park et al., 2008)) were used in this study. They were maintained on irradiated mouse embryonic fibroblasts in hESC medium as previously described (Kennedy et al., 2007). Cells were depleted of feeder cells by culturing on Matrigel (BD Biosciences, Bedford, MA) in hESC medium for 24 to 48 hours prior to differentiation. To generate EBs, hPSCs were treated with collagenase B (1 mg / ml; Roche, Indianapolis, IN) for 20 minutes, followed by a brief trypsin-EDTA (0.05%) step. Gently scrape the cells with a cell scraper to form small aggregates (10-20 cells). The aggregates were resuspended in supplemented with penicillin / streptomycin (10ng / ml), L-glutamine (2mM), ascorbic acid (1mM), thioglycerol (MTG, 4×10 -4 M; Sigma) and transferrin (150 μg / ml) in StemPro-34 (Invitrogen). BMP-4 (1...

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Abstract

There is described herein a method of enriching a population of stem cells for hematopoietic progenitors. The method comprises inducing hematopoietic differentiation in a population of human embryonic stem cells or human induced pluripotent stem cells; sorting the population based on expression of CD43 and at least one of CD34, CD31 and CD144; and selecting a fraction that is at least one of CD34+CD43-, CD31+CD43- and CD144+CD43-. Also provided are populations of hematopoietic progenitors obtained by the methods described herein.

Description

[0001] related application [0002] This application claims priority to US Provisional Patent Application No. 61 / 562,094, filed November 21, 2011, the contents of which are incorporated herein in their entirety. technical field [0003] The present invention relates to populations of hematopoietic progenitor cells and methods of enriching stem cell populations of hematopoietic progenitor cells. Background technique [0004] The ability to generate hematopoietic stem cells (HSCs) from human pluripotent stem cells (PSCs), embryonic (hESC) stem cells, and induced pluripotent stem cells (hiPSCs) will allow the generation of unlimited numbers of patient-matched stem cells for transplantation and derivation studies. New in vitro models of hematopoietic development and disease become possible. Numerous studies have shown that cells of the hematopoietic lineage can be derived from hPSCs by co-culturing them with mesenchymal cells in serum-based media or by directing their different...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0789C12N5/071C12Q1/24
CPCG01N33/56972C12N5/0647C12N2500/24C12N2501/2303C12N2501/2307C12N2501/145C12N2501/16C12N2506/02C12N2501/155C12N2502/1394C12N2501/105C12N2501/26C12N2501/2311C12N2501/14C12N2506/45C12N2500/38C12N2501/165C12N2500/35G01N2333/70596C12N2501/115C12N2501/125C12N2501/2306A61P7/06
Inventor G·凯勒J·C·阻尼加-普鲁科尔M·J·肯尼迪C·M·斯图根A·蒂塔蒂G·S·阿翁
Owner UNIV HEALTH NETWORK
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