在熱敏細胞分選盤上純化人類誘導多能性幹細胞分化的視網膜色素上皮(1/3)

Project Details

Description

There is a lack of organs and tissues for patients in need. Human pluripotent stem cells (hPSCs), which encompass human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), have been considered to be promising sources for regenerating damaged tissues and organs because of their ability to differentiate into cells from three embryonic germ layers. Currently, only for four main diseases are targeted for the clinical trials for stem cell therapy using hPSCs: (1) macular degeneration [age-related macular degeneration (AMD) and Stargardt’s macular dystrophy], (2) acute myocardial infarction (MI), (3) diabetes, and (4) spinal cord injury. The efficient and reproducible differentiation method of hPSCs into retinal pigment epithelium (RPE) has not yet investigated from our database research. Especially, it is unclear what kinds of molecular, physical, and biological design of cell culture biomaterials are preferable for differentiation and purification (sorting) of hPSCs into RPE. Therefore, the development of sophisticated intelligent biomaterials, which are optimized for the differentiation and sorting of hPSCs into RPE are important for future stem cell therapies. In this study, we are going to develop optimal cell culture intelligent biomaterials for differentiation and sorting of hPSCs into RPE in xeno-free conditions. We will investigate (i) what type of intelligent biomaterials are desirable for differentiation of hPSCs into RPE and for sorting of hPSCderived RPE, and (ii) what kind of nanosegments or extracellular matrices (ECMs) should be immobilized on cell culture biomaterials grafted with thermoresponsive polymer (poly(N-isopropylacrylamide), PNIPAAM copolymer) for culture and differentiation of hPSCs as well as sorting of hPSC-derived RPE on intelligent biomaterials. Our originality of the intelligent biomaterial development for stem cell culture, differentiation, and sorting is the combination of physical cues (thermoresponsive polymer-coated dishes to detach specific cells by reducing temperature of the culture medium) and biological cues (selection of specific extracellular matrix (ECM) or ECM-derived oligopeptide for recognition of RPE). We are going to develop cell culture intelligent biomaterials immobilized with thermoresponsive polymer (PNIPAAM copolymer), which are further immobilized with ECM or ECM-derived oligopeptides, which are suitable for stem cell culture, differentiation, and sorting into RPE in clinical usage in future. PNIPAAM copolymer is selected as the thermoresponsive polymer for the coating on the cell culture dishes, which has low critical solution temperature (LCST) around 20 degree. Several ECM proteins and oligopeptides are further immobilized on the PNIPAAM copolymer. The hiPSCs are cultured on the thermoresponsive dishes grafted with ECM and oligopeptides and are differentiated into RPE using several differentiation protocols from articles and from our developed method in this study. We evaluate the efficiency and purity of RPE from the data and discuss the optimal cell culture intelligent biomaterials for specific differentiation and sorting of hPSCs into RPE.
StatusFinished
Effective start/end date1/08/2231/07/23

UN Sustainable Development Goals

In 2015, UN member states agreed to 17 global Sustainable Development Goals (SDGs) to end poverty, protect the planet and ensure prosperity for all. This project contributes towards the following SDG(s):

  • SDG 3 - Good Health and Well-being
  • SDG 17 - Partnerships for the Goals

Keywords

  • intelligent biomaterial
  • extracellular matrix
  • induced pluripotent stem cell
  • retinal pigment epithelium
  • nanosegment
  • differentiation
  • regenerative medicine

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