Methods of mammalian retinal stem cell production and applications

一种哺乳动物、视网膜的技术，应用在哺乳动物视网膜干细胞产生和应用领域，能够解决RPE移植物有效性有限等问题

Pending Publication Date: 2016-07-27
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although hESC-derived RPE transplantation has progressed to clinical trials for the treatment of patients with Stargardt's macular dystrophy and AMD10, RPE transplantation is not an option if most photoreceptors have been lost. may have limited effectiveness

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  • Methods of mammalian retinal stem cell production and applications
  • Methods of mammalian retinal stem cell production and applications
  • Methods of mammalian retinal stem cell production and applications

Examples

Experimental program
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Embodiment 1

[0189] method

[0190] Cell culture and differentiation. Human embryonic stem cells H9 (WA9, WiCell) and HuES9 were cultured in StemProhESCSFM medium (Invitrogen) on plates coated with growth factor-reduced Matrigel (BD Biosciences) under feeder-free and serum-free conditions ( http: / / grants.nih.gov / stem_cells / registry / current.htm? id=40 ) (paragraphs 25-40). according to the previously described procedure 45 Human primary fetal retinal progenitor cells were isolated and cultured from 17-week human fetal retina obtained with informed consent and IRB approval. After undifferentiated hESCs reached approximately 80% confluency in culture, the medium was switched to serum-free N2B27 priming medium (DMEM / F12, N2, B27, 0.2% BSA, 2mML-GlutaMAX, 0.1mMMEM non-essential amino acids and 0.1mM β-mercaptoethanol) for 1-2 days. Near confluent monolayer cultures of hESCs were further grown in N2B27 priming medium supplemented with small molecule inhibitors (5 μM SB431542, 50 nML DN1...

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Abstract

The invention provides an in vitro method for producing isolated mammalian primitive retinal stem cells (pRSCs) comprising: (a) culturing isolated embryonic stem cells (ESCs) from a mammal in a cell culture medium that is free of feeder cells, feeder-conditioned medium or serum so as to produce and grow a culture of the isolated ESCs; and (b) contacting the culture of the isolated ESCs so grown with one or more of an inhibitor for Wnt or TGF-beta / BMP signaling so as to differentiate the isolated ESCs of (a) into primitive retinal stem cells thereby producing isolated mammalian pRSCs.

Description

[0001] This invention was made with government support under grant numbers R01EY021374 and R01EY018660 awarded by the NIH. The government has certain rights in this invention. [0002] Various publications are referenced throughout this application. The disclosures of these publications in their entireties are hereby incorporated by reference into this application to more fully describe the state of the art to which this invention pertains. Background technique [0003] Major causes of irreversible blindness include age-related macular degeneration (AMD), retinitis pigmentosa, glaucoma, and retinal vascular disease that causes RPE, photoreceptors, retinal ganglion cells (RGC) and supporting cells in the retina Loss. One potential treatment is to restore visual function by transplanting healthy retinal cells to replace lost retinal cells. In recent decades, attempts to use primary retinal progenitor cells isolated from human fetal or adult retinal tissue have met with limite...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00C12N5/02
CPCC12N5/0623A61K35/30C12N5/062C12N5/0621C12N2500/33C12N2500/38C12N2501/115C12N2501/15C12N2501/155C12N2501/16C12N2501/165C12N2501/39C12N2501/395C12N2501/415C12N2501/42C12N2501/727C12N2506/02A61L27/52A61K35/545A61L27/24A61L27/26A61L27/3804A61L27/54A61L2300/64A61L2430/16A61P27/02A61P27/06A61K2300/00C08L5/08C08L1/00A61K9/0046A61L27/3834A61L27/3839C12N2500/90C12N2501/999
Inventor K·张J·赵艾达·阿勒穆泰里
Owner RGT UNIV OF CALIFORNIA
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