Procedure of Adult Porcine Islet Isolation

Sakata, Naoaki; Yoshimatsu, Gumpei; Kawakami, Ryo; Chinen, Kiyoshi; Aoyagi, Chikao; Kodama, Shohta

doi:10.1089/ten.tec.2023.0020

Cited by 4 publications

(6 citation statements)

References 59 publications

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“…Procurement of porcine pancreata was described previously 11 , 27 . Total pancreatectomy of donor pigs was performed for organ procurement under general anesthesia using isoflurane (Fujifilm Wako Pure Chemical Co., Osaka, Japan).…”

Section: Methodsmentioning

confidence: 99%

Influence of relatively short-term culture on adult porcine islets for xenotransplantation

Sakata,

Yoshimatsu,

Kawakami

et al. 2024

Sci Rep

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Porcine islet xenotransplantation is a promising therapy for severe diabetes mellitus. Maintenance of the quality and quantity of porcine islets is important for the success of this treatment. Here, we aimed to elucidate the influence of relatively short-term (14 days) culture on adult porcine islets isolated from three micro-minipigs (P111, P112 and P121). Morphological characteristics of islets changed little after 14 days of culture. The viability of cultured islets was also maintained at a high level (> 80%). Furthermore, cultured islets exhibited similar glucose-stimulated insulin secretion and insulin content at Day 14 were preserved comparing with Day 1, while the expressions of Ins, Gcg and Sst were attenuated at Day 14. Xenotransplantation using diabetic nude mice showed no normalization of blood glucose but increased levels of plasma porcine C-peptide after the transplantation of 14 day cultured porcine islets. Histological assessment revealed that relatively short-term cultured porcine islets were successfully engrafted 56 days following transplantation. These data show that relatively short-term culture did not impair the quality of adult porcine islets in regard to function, morphology, and viability. Prevention of impairment of gene correlated with endocrine hormone is warranted for further improvement.

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Section: Methodsmentioning

confidence: 99%

Influence of relatively short-term culture on adult porcine islets for xenotransplantation

Sakata,

Yoshimatsu,

Kawakami

et al. 2024

Sci Rep

Self Cite

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show abstract

“…The porcine pancreas was harvested as described in our previous studies 15 , 37 . In brief, a total pancreatectomy was performed under general anesthesia using isoflurane.…”

Section: Methodsmentioning

confidence: 99%

“…Porcine islet isolation was also conducted as described in our previous studies 15 , 37 . A collagenase solution containing liberase MTF (0.5 g per 1 vial) and thermolysin (15 mg per 1 vial) (Cat# 05339880001; Roche CustomBiotech, Penzberg, Germany) was instilled into the disinfected pancreas via the catheter placed in the pancreatic duct.…”

Section: Methodsmentioning

confidence: 99%

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The porcine islet-derived organoid showed the characteristics as pancreatic duct

Sakata,

Yoshimatsu,

Kawakami

et al. 2024

Sci Rep

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Organoid is a tissue-engineered organ-like structure that resemble as an organ. Porcine islet-derived organoid might be used as an alternative donor of porcine islet xenotransplantation, a promising therapy for severe diabetes. In this study, we elucidated the characteristics of porcine islet organoids derived from porcine islets as a cell source for transplantation. Isolated porcine islets were 3D-cultured using growth factor-reduced matrigel in organoid culture medium consist of advanced DMEM/F12 with Wnt-3A, R-spondin, EGF, Noggin, IGF-1, bFGF, nicotinamide, B27, and some small molecules. Morphological and functional characteristics of islet organoids were evaluated in comparison with 2D-cultured islets in advanced DMEM/F12 medium. Relatively short-term (approximately 14 days)—cultured porcine islet organoids were enlarged and proliferated, but had an attenuated insulin-releasing function. Long-term (over a month)—cultured islet organoids could be passaged and cryopreserved. However, they showed pancreatic duct characteristics, including cystic induction, strong expression of Sox9, loss of PDX1 expression, and no insulin-releasing function. These findings were seen in long-term-cultured porcine islets. In conclusion, our porcine islet organoids showed the characteristics of pancreatic ducts. Further study is necessary for producing porcine islet-derived organoids having characteristics as islets.

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“…For the success of adult porcine islet xenotransplantation, large numbers of high-quality islets must be obtained. However, porcine islet isolation is technically difficult, because of the vulnerability of islets, compared with islet isolation for other species (11,12). Previous studies have shown that the expression of collagen is low in the peripheral regions of porcine islets (13), and the basement membranes of porcine islets are easily damaged during routine islet isolation (14).…”

Section: Introductionmentioning

confidence: 99%

Optimal temperature for the long-term culture of adult porcine islets for xenotransplantation

Sakata,

Yoshimatsu,

Kawakami

et al. 2023

Front. Immunol.

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Porcine islet xenotransplantation represents a promising therapy for severe diabetes mellitus. Long-term culture of porcine islets is a crucial challenge to permit the on-demand provision of islets. We aimed to identify the optimal temperature for the long-term culture of adult porcine islets for xenotransplantation. We evaluated the factors potentially influencing successful 28-day culture of islets at 24°C and 37°C, and found that culture at 37°C contributed to the stability of the morphology of the islets, the proliferation of islet cells, and the recovery of endocrine function, indicated by the expression of genes involved in pancreatic development, hormone production, and glucose-stimulated insulin secretion. These advantages may be provided by islet-derived CD146-positive stellate cells. The efficacy of xenotransplantation using islets cultured for a long time at 37°C was similar to that of overnight-cultured islets. In conclusion, 37°C might be a suitable temperature for the long-term culture of porcine islets, but further modifications will be required for successful xenotransplantation in a clinical setting.

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Procedure of Adult Porcine Islet Isolation

Cited by 4 publications

References 59 publications

Influence of relatively short-term culture on adult porcine islets for xenotransplantation

Influence of relatively short-term culture on adult porcine islets for xenotransplantation

The porcine islet-derived organoid showed the characteristics as pancreatic duct

Optimal temperature for the long-term culture of adult porcine islets for xenotransplantation

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