Cellular models of and therapies for ocular diseases

An eye disease and cell technology, applied in the cell model of eye disease and the field of therapy for eye disease, can solve the problem of lack of good diagnosis of BCD

Pending Publication Date: 2020-09-04
REFLECTION BIOTECH LTD
View PDF20 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, BCD may have been under-diagnosed

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
  • Cellular models of and therapies for ocular diseases
  • Cellular models of and therapies for ocular diseases
  • Cellular models of and therapies for ocular diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0524] Example 1 - Generation and Characterization of Induced Pluripotent Stem Cells (iPSCs) Derived from BCD Patients

[0525] In this study, iPSCs were generated from BCD patients using an integration-free method. Traditional techniques for iPSC reprogramming (eg, lentivirus, retrovirus) integrate into the genome of the target cell. The resulting iPSCs and cells differentiated from those iPSCs will contain exogenous DNA and may be unsafe and potentially problematic for use in cell therapy and drug discovery applications. Furthermore, integration may occur in critical regions of the genome, causing problems in unrelated developmental processes. Compared to traditional reprogramming methods, integration-free reprogramming methods generate iPSCs without detectable vectors or transgenes, making them more suitable for cell therapy and drug discovery applications.

[0526] In this study, iPSCs were generated from BCD patients using two different integration-free reprogramming ...

Embodiment 2

[0548] Example 2 - Differentiation of iPSCs from BCD patients into retinal pigment epithelial (RPE) cells

[0549]The iPSC differentiation of all iPSC lines from BCD patients and healthy controls started from passage 3 to passage 6. For differentiation, iPS colonies were grown to confluence for the first 14 days in differentiation medium in 6-well dishes (Costar, Corning, Corning, NY) pretreated with Matrigel (CORNING, 356230) diluted 1:50 , the differentiation medium consisted of knockout (KO) DMEM (Thermo Fisher Scientific, 10829018), 15% KO serum substitute (Thermo Fisher Scientific, 10829028), 1% non-essential amino acids (Thermo Fisher Scientific, 11140050), 2 mmol / L gluten Aminoamide (Thermo Fisher Scientific, 35050061), 50 U / ml penicillin-streptomycin (ThermoFisher Scientific, 10378016) and 10 mmol / L nicotinamide (Sigma-Aldrich, N0636). During the 15th to 28th day of differentiation, the differentiation medium was supplemented with 100 ng / ml human activin-A (PeproTec...

Embodiment 3

[0552] Biochemical analysis, cell viability analysis and other analysis of embodiment 3-BCD cell model and CYP4V2 function research study

[0553] Lipid Analysis:

[0554] Previous studies on the function of BCD and CYP4V2 enzymes have focused on fatty acids. In this study, more lipid assays including not only fatty acids but also ceramide (Cer), sphingomyelin, (SM) and sphingosine and dihydrosphingosine (SOSA).

[0555] The biochemical analysis of free fatty acid (FFA), ceramide (Cer), sphingomyelin (SM), sphingosine and dihydrosphingosine (SOSA) was carried out at Columbia University (New York, NY). , USA) based on its relevant analysis and protocols.

[0556] Free fatty acids (FFA), ceramides, sphingosines and dihydrosphingosines were extracted by using chloroform:methanol. Briefly, approximately 1 million iPS-RPE cells were homogenized in 150 µL of water. Mix 100 μL of homogenate with 3 mL of chloroform:methanol (v:v=2:1) ​​containing internal standard (palmitic a...

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

Cellular models for diseases of eyes, methods and compositions for treating diseases in eyes are provided.

Description

[0001] Cross References to Related Applications [0002] This application claims priority under 35 U.S.C. §119(e) to U.S. Application No. 62 / 539,473, filed July 31, 2017, entitled "CELLULARMODELS OF AND THERAPIES FOR OCULAR DISEASES." The entire content of the above application is hereby incorporated by reference. 【Background technique】 [0003] Background of the invention [0004] Crystalline retinitis pigmentosa (also called crystalline retinal degeneration, or Bietti's Crystalline Dystrophy, BCD) [0005] Crystalline retinitis pigmentosa (BCD, also known as Bietti Crystalline Corneoretinal Dystrophy, Bietti Crystalline Retinopathy, Bietti's Retinal Dystrophy (OMIM 210370)) is a rare and often Chromosomal recessive and blinding retinal degeneration characterized by the presence of numerous fine, shiny yellow-white crystal-like deposits at the posterior pole of the retina, associated with atrophy of the retinal pigment epithelium (RPE), pigment clumps ) and choroidal scler...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/63C12N5/22A61K48/00A61P27/02
CPCA61P27/02C12N2750/14143A61K48/0075A61K48/005A61K38/1709C07K14/80G01N33/5073C12N15/102C12N15/113C12N2310/20C12N9/22C12N15/86
Inventor 杨瑞曾庆霖
Owner REFLECTION BIOTECH LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap