In vitro and in vivo effects of insulin-producing cells generated by xeno-antigen free 3D culture with RCP piece

Abstract To establish widespread cell therapy for type 1 diabetes mellitus, we aimed to develop an effective protocol for generating insulin-producing cells (IPCs) from adipose-derived stem cells (ADSCs). We established a 3D culture using a human recombinant peptide (RCP) petaloid μ-piece with xeno-...

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Autores principales: Tetsuya Ikemoto, Rui Feng, Shu-ichi Iwahashi, Shinichiro Yamada, Yu Saito, Yuji Morine, Satoru Imura, Munehide Matsuhisa, Mitsuo Shimada
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Publicado: Nature Portfolio 2019
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spelling oai:doaj.org-article:d40d9f4dd7634f25b2ef7f90de61850e2021-12-02T15:09:38ZIn vitro and in vivo effects of insulin-producing cells generated by xeno-antigen free 3D culture with RCP piece10.1038/s41598-019-47257-72045-2322https://doaj.org/article/d40d9f4dd7634f25b2ef7f90de61850e2019-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-47257-7https://doaj.org/toc/2045-2322Abstract To establish widespread cell therapy for type 1 diabetes mellitus, we aimed to develop an effective protocol for generating insulin-producing cells (IPCs) from adipose-derived stem cells (ADSCs). We established a 3D culture using a human recombinant peptide (RCP) petaloid μ-piece with xeno-antigen free reagents. Briefly, we employed our two-step protocol to differentiate ADSCs in 96-well dishes and cultured cells in xeno-antigen free reagents with 0.1 mg/mL RCP μ-piece for 7 days (step 1), followed by addition of histone deacetylase inhibitor for 14 days (step 2). Generated IPCs were strongly stained with dithizone, anti-insulin antibody at day 21, and microstructures resembling insulin secretory granules were detected by electron microscopy. Glucose stimulation index (maximum value, 4.9) and MAFA mRNA expression were significantly higher in 3D cultured cells compared with conventionally cultured cells (P < 0.01 and P < 0.05, respectively). The hyperglycaemic state of streptozotocin-induced diabetic nude mice converted to normoglycaemic state around 14 days after transplantation of 96 IPCs under kidney capsule or intra-mesentery. Histological evaluation revealed that insulin and C-peptide positive structures existed at day 120. Our established xeno-antigen free and RCP petaloid μ-piece 3D culture method for generating IPCs may be suitable for clinical application, due to the proven effectiveness in vitro and in vivo.Tetsuya IkemotoRui FengShu-ichi IwahashiShinichiro YamadaYu SaitoYuji MorineSatoru ImuraMunehide MatsuhisaMitsuo ShimadaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-9 (2019)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Tetsuya Ikemoto
Rui Feng
Shu-ichi Iwahashi
Shinichiro Yamada
Yu Saito
Yuji Morine
Satoru Imura
Munehide Matsuhisa
Mitsuo Shimada
In vitro and in vivo effects of insulin-producing cells generated by xeno-antigen free 3D culture with RCP piece
description Abstract To establish widespread cell therapy for type 1 diabetes mellitus, we aimed to develop an effective protocol for generating insulin-producing cells (IPCs) from adipose-derived stem cells (ADSCs). We established a 3D culture using a human recombinant peptide (RCP) petaloid μ-piece with xeno-antigen free reagents. Briefly, we employed our two-step protocol to differentiate ADSCs in 96-well dishes and cultured cells in xeno-antigen free reagents with 0.1 mg/mL RCP μ-piece for 7 days (step 1), followed by addition of histone deacetylase inhibitor for 14 days (step 2). Generated IPCs were strongly stained with dithizone, anti-insulin antibody at day 21, and microstructures resembling insulin secretory granules were detected by electron microscopy. Glucose stimulation index (maximum value, 4.9) and MAFA mRNA expression were significantly higher in 3D cultured cells compared with conventionally cultured cells (P < 0.01 and P < 0.05, respectively). The hyperglycaemic state of streptozotocin-induced diabetic nude mice converted to normoglycaemic state around 14 days after transplantation of 96 IPCs under kidney capsule or intra-mesentery. Histological evaluation revealed that insulin and C-peptide positive structures existed at day 120. Our established xeno-antigen free and RCP petaloid μ-piece 3D culture method for generating IPCs may be suitable for clinical application, due to the proven effectiveness in vitro and in vivo.
format article
author Tetsuya Ikemoto
Rui Feng
Shu-ichi Iwahashi
Shinichiro Yamada
Yu Saito
Yuji Morine
Satoru Imura
Munehide Matsuhisa
Mitsuo Shimada
author_facet Tetsuya Ikemoto
Rui Feng
Shu-ichi Iwahashi
Shinichiro Yamada
Yu Saito
Yuji Morine
Satoru Imura
Munehide Matsuhisa
Mitsuo Shimada
author_sort Tetsuya Ikemoto
title In vitro and in vivo effects of insulin-producing cells generated by xeno-antigen free 3D culture with RCP piece
title_short In vitro and in vivo effects of insulin-producing cells generated by xeno-antigen free 3D culture with RCP piece
title_full In vitro and in vivo effects of insulin-producing cells generated by xeno-antigen free 3D culture with RCP piece
title_fullStr In vitro and in vivo effects of insulin-producing cells generated by xeno-antigen free 3D culture with RCP piece
title_full_unstemmed In vitro and in vivo effects of insulin-producing cells generated by xeno-antigen free 3D culture with RCP piece
title_sort in vitro and in vivo effects of insulin-producing cells generated by xeno-antigen free 3d culture with rcp piece
publisher Nature Portfolio
publishDate 2019
url https://doaj.org/article/d40d9f4dd7634f25b2ef7f90de61850e
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