Hisato Nagano

All Publications

• An intrinsically semi-permeable PDMS nanosheet encapsulating adipose tissue-derived stem cells for enhanced angiogenesis. Biomaterials science Takuma, M., Fujita, H., Zushi, N., Nagano, H., Azuma, R., Kiyosawa, T., Fujie, T. 2024

  Abstract

  Cell encapsulation devices are expected to be promising tools that can control the release of therapeutic proteins secreted from transplanted cells. The protein permeability of the device membrane is important because it allows the isolation of transplanted cells while enabling the effectiveness of the device. In this study, we investigated free-standing polymeric ultra-thin films (nanosheets) as an intrinsically semi-permeable membrane made from polydimethylsiloxane (PDMS). The PDMS nanosheet with a thickness of 600 nm showed intrinsic protein permeability, and the device fabricated with the PDMS nanosheet showed that VEGF secreted from implanted adipose tissue-derived stem cells (ASCs) could be released for at least 5 days. The ASC encapsulation device promoted angiogenesis and the development of granulation tissue 1 week after transplantation to the subcutaneous area of a mouse. This cell encapsulation device consisting of PDMS nanosheets provides a new method for pre-vascularization of the subcutaneous area in cell transplantation therapy.

  View details for DOI 10.1039/d4bm00460d

  View details for PubMedID 38804980

• Skin graft with dermis and appendages generated in vivo by cell competition. Nature communications Nagano, H., Mizuno, N., Sato, H., Mizutani, E., Yanagida, A., Kano, M., Kasai, M., Yamamoto, H., Watanabe, M., Suchy, F., Masaki, H., Nakauchi, H. 2024; 15 (1): 3366

  Abstract

  Autologous skin grafting is a standard treatment for skin defects such as burns. No artificial skin substitutes are functionally equivalent to autologous skin grafts. The cultured epidermis lacks the dermis and does not engraft deep wounds. Although reconstituted skin, which consists of cultured epidermal cells on a synthetic dermal substitute, can engraft deep wounds, it requires the wound bed to be well-vascularized and lacks skin appendages. In this study, we successfully generate complete skin grafts with pluripotent stem cell-derived epidermis with appendages on p63 knockout embryos' dermis. Donor pluripotent stem cell-derived keratinocytes encroach the embryos' dermis by eliminating p63 knockout keratinocytes based on cell-extracellular matrix adhesion mediated cell competition. Although the chimeric skin contains allogenic dermis, it is engraftable as long as autologous grafts. Furthermore, we could generate semi-humanized skin segments by human keratinocytes injection into the amnionic cavity of p63 knockout mice embryos. Niche encroachment opens the possibility of human skin graft production in livestock animals.

  View details for DOI 10.1038/s41467-024-47527-7

  View details for PubMedID 38684678

  View details for PubMedCentralID PMC11058811

• Functional calcium-responsive parathyroid glands generated using single-step blastocyst complementation. Proceedings of the National Academy of Sciences of the United States of America Kano, M., Mizuno, N., Sato, H., Kimura, T., Hirochika, R., Iwasaki, Y., Inoshita, N., Nagano, H., Kasai, M., Yamamoto, H., Yamaguchi, T., Suga, H., Masaki, H., Mizutani, E., Nakauchi, H. 2023; 120 (28): e2216564120

  Abstract

  Patients with permanent hypoparathyroidism require lifelong replacement therapy to avoid life-threatening complications, The benefits of conventional treatment are limited, however. Transplanting a functional parathyroid gland (PTG) would yield better results. Parathyroid gland cells generated from pluripotent stem cells in vitro to date cannot mimic the physiological responses to extracellular calcium that are essential for calcium homeostasis. We thus hypothesized that blastocyst complementation (BC) could be a better strategy for generating functional PTG cells and compensating loss of parathyroid function. We here describe generation of fully functional PTGs from mouse embryonic stem cells (mESCs) with single-step BC. Using CRISPR-Cas9 knockout of Glial cells missing2 (Gcm2), we efficiently produced aparathyroid embryos for BC. In these embryos, mESCs differentiated into endocrinologically mature PTGs that rescued Gcm2-/- mice from neonatal death. The mESC-derived PTGs responded to extracellular calcium, restoring calcium homeostasis on transplantation into mice surgically rendered hypoparathyroid. We also successfully generated functional interspecies PTGs in Gcm2-/- rat neonates, an accomplishment with potential for future human PTG therapy using xenogeneic animal BC. Our results demonstrate that BC can produce functional endocrine organs and constitute a concept in treatment of hypoparathyroidism.

  View details for DOI 10.1073/pnas.2216564120

  View details for PubMedID 37379351