Poursafavi Zahra, Abroun Saeid, Kaviani Saeid, Hayati Roodbari Nasim
Biology Department, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.
Cell J. 2022 Aug 28;24(8):449-457. doi: 10.22074/cellj.2022.8081.
Objective: Insulin insufficiency due to the reduced pancreatic beta cell number is the hallmark of diabetes, resulting in
an intense focus on the development of beta-cell replacement options. One approach to overcome the problem is to
search for alternative sources to induce insulin-producing cells (IPCs), the advent of mesenchymal stem cells (MSCs)
holds great promise for producing ample IPCs. Encapsulate the MSCs with alginate improved anti-inflammatory effects
of MSC treatment. This study aimed to evaluate the differentiation of wharton jelly-derived MScs into insulin producing
cells using alginate encapsulation.
Materials and Methods: In this experimental study, we established an efficient IPCs differentiation strategy of human
MSCs derived from the umbilical cord's Wharton jelly with lentiviral transduction of Pancreas/duodenum homeobox
protein 1 (PDX1) in a 21-day period using alginate encapsulation by poly-L-lysine (PLL) and poly-L-ornithine (PLO)
outer layer. During differentiation, the expression level of PDX1 and secretion of insulin proteins were increased.
Results: Results showed that during time, the cell viability remained high at 87% at day 7. After 21 days, the differentiated beta-like cells in microcapsules were morphologically similar to primary beta cells. Evaluation of the expression of PDX1 and INS by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) on days 7, 14 and 21 of differentiation exhibited the highest expression on day 14. At the protein level, the expression of these two pancreatic markers was observed after PDX1 transduction. Results showed that the intracellular and extracellular insulin levels in the cells receiving PDX1 is higher than the control group. The current data showed that encapsulation with alginate by PLL and PLO outer layer permitted to increase the microcapsules' beta cell differentiation.
Conclusion: Encapsulate the transduced-MSCs with alginate can be applied in an in vivo model in order to do the further analysis.
目的：胰腺β细胞数量减少导致的胰岛素分泌不足是糖尿病的标志，这使得人们将重点高度集中在β细胞替代方案的开发上。解决该问题的一种方法是寻找替代来源来诱导产生胰岛素的细胞（IPC），间充质干细胞（MSC）的出现为大量产生IPC带来了巨大希望。用海藻酸盐包裹MSC可改善MSC治疗的抗炎效果。本研究旨在评估使用海藻酸盐包裹将脐带来源的华通胶间充质干细胞分化为产生胰岛素的细胞。
材料和方法：在本实验研究中，我们建立了一种高效的人脐带华通胶间充质干细胞向IPC分化的策略，通过胰腺/十二指肠同源盒蛋白1（PDX1）的慢病毒转导，在21天内使用聚-L-赖氨酸（PLL）和聚-L-鸟氨酸（PLO）外层进行海藻酸盐包裹。在分化过程中，PDX1的表达水平和胰岛素蛋白的分泌增加。
结果：结果显示，在第7天时细胞活力保持在较高水平，为87%。21天后，微胶囊中分化的β样细胞在形态上与原代β细胞相似。在分化的第7天、14天和21天，通过定量逆转录聚合酶链反应（qRT-PCR）评估PDX1和胰岛素（INS）的表达，结果显示在第14天表达最高。在蛋白质水平上，在PDX1转导后观察到这两种胰腺标志物的表达。结果表明，接受PDX1转导的细胞内和细胞外胰岛素水平均高于对照组。目前的数据表明，用PLL和PLO外层的海藻酸盐包裹可促进微胶囊中β细胞的分化。
结论：用海藻酸盐包裹转导后的间充质干细胞可应用于体内模型以进行进一步分析。
