Introduction

Developing transplantable humanized endocrine grafts for long-term management of disorders like uncontrolled diabetes mellitus has been one of the most challenging concerns in tissue and organ bioengineering. Hence, we aimed to develop bio/immune compatible and transplantable humanized insulin producing neo-organs through biofabrication of rat spleen which acts as secondary neo-organoid after ectopic transplantation.

Methods

An efficient process was developed using splenic artery perfusion to generate completely cell deficient decellularized splenic matrices (DSM). Microscopic, vascular tree imaging, mechanical strength and other required analysis were performed before and after decellularization. Biofabrication of humanized insulin producing neo-organ was performed through infusion of hyperglycaemic challenged trans-differentiated human hepatic progenitor cells (hHPCs) in DSM. Intra-peritoneal transplantation of biofabricated insulin producing humanized neo-organs in rats was done to evaluate the bio/immune compatibility.

Results

The biofabrication strategy generated highly intact DSM with preserved organ architecture, extracellular matrix and organ vasculature. Repopulation of DSM with glucose responsive cells showed enhanced engraftment efficiency, survival and proliferation. Bioengineered humanized insulin producing neo-organs showed higher amount of insulin secretion than conventional 2D-cultured cells in response to hyperglycaemic stimuli ex vivo. Glucose responsive humanized neo-organs survived in vivo without eliciting immunological or fibrotic responses and proved to be secondary neo-organoids at ectopic site with the development of neo-vasculature and responded to hyperglycaemic challenge.

Conclusion

The present study provides first proof-of-concept for biofabricating bio/immune compatible, humanized insulin producing neo-organoids post-ectopic transplantation which provides more suitable functional biological implants for long-term management of uncontrolled diabetes mellitus.

中文翻译：

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生物制造人源化的胰岛素生产新器官，通过异位移植产生次生新器官。

介绍

开发可移植的人源化内分泌移植物以长期治疗诸如失控​​的糖尿病等疾病一直是组织和器官生物工程领域最具挑战性的问题之一。因此，我们旨在通过大鼠脾脏的生物制造来开发生物/免疫相容且可移植的人源化胰岛素生产新器官，所述大鼠脾脏在异位移植后充当继发性新有机体。

方法

开发了一种有效的方法，可以使用脾动脉灌注来生成完全细胞缺陷的脱细胞脾脏基质（DSM）。在脱细胞之前和之后进行显微镜，血管树成像，机械强度和其他所需的分析。通过在DSM中输注高血糖攻击的反分化人类肝祖细胞（hHPC），进行人源化胰岛素生产新生物的生物制造。进行了大鼠体内产生人源化新器官的生物合成胰岛素的腹膜内移植，以评估其生物/免疫相容性。

结果

生物制造策略生成了完整的DSM，并保留了器官结构，细胞外基质和器官脉管系统。用葡萄糖反应性细胞重新补充DSM表现出更高的植入效率，存活和增殖。生物工程化的人源化产生胰岛素的新器官对离体高血糖刺激的反应显示出比传统的2D培养细胞更高的胰岛素分泌量。葡萄糖反应性人源化新器官在体内存活而未引起免疫学或纤维化反应，并且随着新脉管系统的发展而被证明是异位部位的继发性新有机体，并对高血糖挑战做出了反应。

结论

本研究提供了生物制造生物/免疫相容的，人源化的胰岛素生产新有机体的异位移植后的第一个概念验证，这为长期控制不受控制的糖尿病提供了更合适的功能性生物植入物。