CellSaic, A Cell Aggregate-Like Technology Using Recombinant Peptide Pieces for MSC Transplantation

Nakamura, Kentaro

doi:10.2174/1574888x13666180912125157

Cited by 17 publications

(17 citation statements)

References 35 publications

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“…The advantage of μ -pieces, such as human recombinant peptide petaloid, was reported for islet cell culture and cell transplantation [ 41 ]. Spheroid-like cell aggregation-formed combination cells with μ -pieces can prevent central necrosis within cell aggregates, because nutrients and waste products can pass through the inner spheroid [ 42 ]. Our results showed that IPC generated from canine AT-MSCs in the 3D culture system showed better DTZ staining and insulin secretion compared with cells in conventional 2D culture dishes.…”

Section: Discussionmentioning

confidence: 99%

Generation of Insulin-Producing Cells from Canine Adipose Tissue-Derived Mesenchymal Stem Cells

Teshima

Okamoto

Dairaku

et al. 2020

Stem Cells International

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The potential of mesenchymal stem cells (MSCs) to differentiate into nonmesodermal cells such as pancreatic beta cells has been reported. New cell-based therapy using MSCs for diabetes mellitus is anticipated as an alternative treatment option to insulin injection or islet transplantation in both human and veterinary medicine. Several protocols were reported for differentiation of MSCs into insulin-producing cells (IPCs), but no studies have reported IPCs generated from canine MSCs. The purpose of this study was to generate IPCs from canine adipose tissue-derived MSCs (AT-MSCs) in vitro and to investigate the effects of IPC transplantation on diabetic mice in vivo. Culturing AT-MSCs with the differentiation protocol under a two-dimensional culture system did not produce IPCs. However, spheroid-like small clusters consisting of canine AT-MSCs and human recombinant peptide μ-pieces developed under a three-dimensional (3D) culture system were successfully differentiated into IPCs. The generated IPCs under 3D culture condition were stained with dithizone and anti-insulin antibody. Canine IPCs also showed gene expression typical for pancreatic beta cells and increased insulin secretion in response to glucose stimulation. The blood glucose levels in streptozotocin-induced diabetic mice were decreased after injection with the supernatant of canine IPCs, but the hyperglycemic states of diabetic mice were not improved after transplanting IPCs subcutaneously or intramesenterically. The histological examination showed that the transplanted small clusters of IPCs were successfully engrafted to the mice and included cells positive for insulin by immunofluorescence. Several factors, such as the transplanted cell number, the origin of AT-MSCs, and the differentiation protocol, were considered potential reasons for the inability to improve the hyperglycemic state after IPC transplantation. These findings suggest that canine AT-MSCs can be differentiated into IPCs under a 3D culture system and IPC transplantation may be a new treatment option for dogs with diabetes mellitus.

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Section: Discussionmentioning

confidence: 99%

Generation of Insulin-Producing Cells from Canine Adipose Tissue-Derived Mesenchymal Stem Cells

Teshima

Okamoto

Dairaku

et al. 2020

Stem Cells International

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“…Here we developed a 3D culture system with a RCP petaloid μ-piece to successfully generate effective IPCs that marked 65.6% of cell expansion rate (according to DNA quantity, data not shown) and reasonable SI value. Moreover, this RCP petaloid μ-piece is xeno-antigen free and is provided as clinical grade 22,23 . Our established two-step generation protocol is thus a xeno-antigen-free protocol.…”

Section: Discussionmentioning

confidence: 99%

“…The CellSaic maintains cell viability in vivo when transplanted with target cells. Additionally, CellSaic is xeno-antigen free and clinical grade has been achieved 23 .…”

Section: Introductionmentioning

confidence: 99%

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

Ikemoto

Feng

Iwahashi

et al. 2019

Sci Rep

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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 .

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“…Thus, the focus of this study is on the ADSCs as a new cell source of MSCs 6–8 and the establishment of a new 2-step protocol for the differentiation of IPCs from ADSCs with a short culture duration and functional efficiency 9 . Moreover, this 2-step protocol was modified into a xeno-antigen free and three-dimensional (3D) culture protocol 10,11 which improved cell quality and function in vitro / in vivo and resulted in the normalization of diabetic nude mice blood glucose over 120 days 12 . In other words, this modified protocol is reaching the pre-clinical level of proof of a concept.…”

Section: Introductionmentioning

confidence: 99%

The Differences in the Characteristics of Insulin-producing Cells Using Human Adipose-tissue Derived Mesenchymal Stem Cells from Subcutaneous and Visceral Tissues

Wada

Ikemoto

Morine

et al. 2019

Sci Rep

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The aim of this study was to investigate the characteristics of insulin producing cells (IPCs) differentiated from adipose-tissue derived stem cells (ADSCs) isolated from human subcutaneous and visceral adipose tissues and identify ADSCs suitable for differentiation into efficient and functional IPCs. Subcutaneous and visceral adipose tissues collected from four (4) patients who underwent digestive surgeries at The Tokushima University (000035546) were included in this study. The insulin secretion of the generated IPCs was investigated using surface markers by: fluorescence activated cell sorting (FACS) analysis; cytokine release; proliferation ability of ADSCs; in vitro (glucose-stimulated insulin secretion: (GSIS) test/in vivo (transplantation into streptozotocin-induced diabetic nude mice). The less fat-related inflammatory cytokines secretions were observed (P < 0.05), and the proliferation ability was higher in the subcutaneous ADSCs (P < 0.05). Insulin expression and GISI were higher in the subcutaneous IPCs (P < 0.01 and P < 0.05, respectively). The hyperglycaemic state of all mice that received IPCs from subcutaneous fat tissue converted into normo-glycaemia in thirty (30) days post-transplantation (4/4,100%). Transplanted IPCs were stained using anti-insulin and anti-human leukocyte antigen antibodies. The IPCs generated from the ADSCs freshly isolated from the human fat tissue had sufficient insulin secreting ability in vitro and in vivo.

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CellSaic, A Cell Aggregate-Like Technology Using Recombinant Peptide Pieces for MSC Transplantation

Cited by 17 publications

References 35 publications

Generation of Insulin-Producing Cells from Canine Adipose Tissue-Derived Mesenchymal Stem Cells

Generation of Insulin-Producing Cells from Canine Adipose Tissue-Derived Mesenchymal Stem Cells

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

The Differences in the Characteristics of Insulin-producing Cells Using Human Adipose-tissue Derived Mesenchymal Stem Cells from Subcutaneous and Visceral Tissues

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