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体外人胎儿胰岛重新定义胰腺研究。

In Vitro Human Fetal Pancreatic Islets to Redefine Pancreatic Research.

作者信息

Rout Sipra, Amirtham Soosai Manickam, Prasad Mythraeyee, Cherian Anne George, B Sandya Rani, Sudhakar Yesudas, Prince Neetu

机构信息

Anatomy, All India Institute of Medical Sciences, Bhubaneswar, IND.

Physiology, Christian Medical College and Hospital, Vellore, IND.

出版信息

Cureus. 2023 Aug 9;15(8):e43244. doi: 10.7759/cureus.43244. eCollection 2023 Aug.

Abstract

BACKGROUND

In vitro studies with human fetal islets of different gestational ages (GA) would be a great tool to generate information on the developmental process of the islets as this would help to recontextualize diabetes research and clinical practice. Pancreatic islets from human cadavers and other animal species are extensively researched to explore their suitability for islet transplantation procedure, one of the upcoming treatment strategies for insulin-dependent diabetes mellitus. Although human fetal islets are also considered for islet transplantation, ethical issues and limited knowledge constraints their use. The fetal islets could be explored to address the information lacunae on the maturity process of pancreatic islets and the endocrine-exocrine signaling mechanisms.

AIM

This study aimed to assess the feasibility of isolating viable islets and study the cytoarchitecture of the fetal pancreas of GA 22-29 weeks, not reported otherwise.

METHODOLOGY

Pancreas obtained from the aborted fetuses of GA 22-29 weeks were subjected to collagenase digestion and were further cultured to determine the viability in vitro. Parameters assessed were expression of markers for endocrine cell lineages and insulin release to glucose challenge.

RESULTS

Islets were viable in vitro and islets were shown to maintain cues for post-digestion re-aggregation and expansion in culture. The immunofluorescent staining showed islets of varying sizes, homogenous cell clusters aggregating to form heterogenous cell clusters, otherwise not reported for this GA. On stimulation with different concentrations of glucose (2.8 and 28 mM), the fetal islets in the culture exhibited insulin release, and this response confirmed their viability in vitro.

CONCLUSION

Our findings showed that viable islets could be isolated and cultured in vitro for further in-depth studies to explore their proliferative potential as well as for the identification of pancreatic progenitors, a good strategy to take forward.

摘要

背景

利用不同胎龄(GA)的人类胎儿胰岛进行体外研究,将是获取胰岛发育过程信息的有力工具,因为这有助于重新定位糖尿病研究和临床实践。人们广泛研究来自人类尸体和其他动物物种的胰岛,以探索它们是否适合胰岛移植手术,这是胰岛素依赖型糖尿病即将采用的治疗策略之一。尽管人类胎儿胰岛也被考虑用于胰岛移植,但伦理问题和知识有限限制了它们的使用。可以探索利用胎儿胰岛来填补胰岛成熟过程以及内分泌-外分泌信号机制方面的信息空白。

目的

本研究旨在评估分离有活力胰岛的可行性,并研究22 - 29周胎龄胎儿胰腺的细胞结构,此前未见相关报道。

方法

对取自22 - 29周堕胎胎儿的胰腺进行胶原酶消化,并进一步培养以确定其体外活力。评估的参数包括内分泌细胞谱系标志物的表达以及对葡萄糖刺激的胰岛素释放情况。

结果

胰岛在体外具有活力,并且显示出在消化后重新聚集和在培养中扩增的迹象。免疫荧光染色显示大小各异的胰岛,同质细胞簇聚集形成异质细胞簇,在该胎龄下此前未见此报道。在用不同浓度葡萄糖(2.8和28 mM)刺激后,培养中的胎儿胰岛表现出胰岛素释放,这一反应证实了它们在体外的活力。

结论

我们的研究结果表明,可以分离有活力的胰岛并在体外进行培养,以进一步深入研究其增殖潜力以及鉴定胰腺祖细胞,这是一个值得推进的良好策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fac/10491859/eeee723eca75/cureus-0015-00000043244-i01.jpg

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