開發細胞分選培養材料:純化自hiPSC分化的心肌細胞

Project Details

Description

Stem cells, especially human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs) are attractive cell source for regenerative medicine and these stem cells have been used in clinical trials and usages, although adult stem cells such as bone marrow stem cells, umbilical cord blood stem cells and adipose-derived stem cells are more frequently used for the treatment of variety of diseases including graft-versus host disease. One of the difficulties of application of human pluripotent stem cells, hPSCs (hESCs and hiPSCs) into clinics is that it is difficult to differentiate hPSCs with high purity of the differentiated cells without hPSCs. Recently, it is known that physical cues and biochemical cues of cell culture biomaterials guide stem cell fate of differentiation. Currently, 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 specific lineage of the cells such as cardiac progenitors andcardiomyocytes. Therefore, the development of sophisticated intelligent biomaterials, which are optimized for the differentiation and sorting of hPSCs into cardiomyocytes and cardiac progenitors are important for future stem celltherapies. In this study, we are going to develop optimal cell culture intelligent biomaterials for differentiation and sorting of hPSCs into cardiac progenitors and cardiomyocytes in xeno-free conditions. We will investigate (i) what type of intelligent biomaterials are desirable for differentiation of hPSCs into cardiomyocyte progenitors and cardiomyocytes and for sorting of hPSC-derived cardiomyocyte progenitors and cardiomyocytes, and (ii) what kind of nanosegments or extracellular matrices (ECMs) should be immobilized on cell culture biomaterials immobilized with thermoresponsive polymer (poly(Nisopropylacrylamide), PNIPAAM, copolymer for culture and differentiation ofhPSCs as well as sorting of hPSC-derived cardiac progenitors and cardiomyocytes 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-immobilized dishes todetach 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 cardiac progenitors and cardiomyocytes). 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 cardiac progenitors and cardiomyocytes in clinical usage in future. PNIPAAM copolymer is selected as the thermoresponsive polymer for the immobilization on the cell culture dishes, which has low criticalsolution temperature (LCST) around 20 degree. Several ECM proteins and oligopeptides are further immobilized on the PNIPAAM copolymer surface. The hiPSCs are cultured on the thermoresponsive dishes grafted with ECM andoligopeptides and are differentiated into cardiac progenitors and cardiomyocytes using several differentiation protocols from articles and from our developed method in this study. We evaluate the efficiency and purity of cardiomyocyte differentiation from the results and discuss the optimal cell culture intelligent biomaterials for specific differentiation and sorting of hPSCs into cardiac progenitors and cardiomyocytes.
StatusFinished
Effective start/end date1/08/2131/07/22

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

Keywords

  • Biomaterial
  • Peptide
  • Cell sorting
  • Extracellular matrix
  • Induced pluripotent stem cell
  • Cardiomyocyte
  • Elasticity
  • Nanosegment
  • Differentiation
  • Regenerative medicine

Fingerprint

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  • Universal and hypoimmunogenic pluripotent stem cells for clinical usage

    Sung, T. C., Maitiruze, K., Pan, J., Gong, J., Bai, Y., Pan, X. & Higuchi, A., Jan 2023, Progress in Molecular Biology and Translational Science. Higuchi, A., Zhou, Y. & Chiou, S.-H. (eds.). Elsevier B.V., p. 271-296 26 p. (Progress in Molecular Biology and Translational Science; vol. 199).

    Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

    1 Scopus citations
  • Xeno-free culture and proliferation of hPSCs on 2D biomaterials

    Wang, T., Yu, T., Tsai, C. Y., Hong, Z. Y., Chao, W. H., Su, Y. S., Subbiah, S. K., Renuka, R. R., Hsu, S. T., Wu, G. J. & Higuchi, A., Jan 2023, Progress in Molecular Biology and Translational Science. Higuchi, A., Zhou, Y. & Chiou, S.-H. (eds.). Elsevier B.V., p. 63-107 45 p. (Progress in Molecular Biology and Translational Science; vol. 199).

    Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

  • Purification of colon carcinoma cells from primary colon tumor using a filtration method via porous polymeric filters

    Wang, J. H., Ban, L. K., Lee, H. H. C., Chen, Y. H., Lin, H. Y., Zhu, Z. W., Su, H. Y., Umezawa, A., Almansour, A. I., Arumugam, N., Kumar, R. S., Wu, G. J. & Higuchi, A., 1 Oct 2021, In: Polymers. 13, 19, 3411.

    Research output: Contribution to journalArticlepeer-review

    Open Access