Colella Antonella, Biondi Giuseppina, Marrano Nicola, Francioso Edda, Fracassi Laura, Crovace Alberto M, Recchia Alessandra, Natalicchio Annalisa, Paradies Paola
Department of Precision and Regenerative Medicine and Ionian Area (DiMePre-J), Veterinary Clinics and Animal Production Section, University of Bari Aldo Moro, Valenzano, 70010 Bari, Italy.
Department of Precision and Regenerative Medicine and Ionian Area (DiMePre-J), Internal Medicine, Endocrinology, Andrology and Metabolic Diseases Section, University of Bari Aldo Moro, 70124 Bari, Italy.
Vet Sci. 2024 Aug 18;11(8):380. doi: 10.3390/vetsci11080380.
Cell-based therapy using insulin-producing cells (IPCs) is anticipated as an alternative treatment option to insulin injection or pancreatic islet transplantation for the treatment of diabetes mellitus in both human and veterinary medicine. Several protocols were reported for the differentiation of mesenchymal stem cells (MSCs) into IPCs; to date, glucose-responsive IPCs have only been obtained from canine adipose tissue-derived MSCs (c-MSCs), but not from canine bone marrow-derived MSCs (cBM-MSCs). Therefore, this study aims to generate in vitro glucose-responsive IPCs from cBM-MSCs using two differentiation protocols: a two-step protocol using trichostatin (TSA) and a three-step protocol using mercaptoethanol to induce pancreatic and duodenal homeobox gene 1 (PDX-1) expression. A single experiment was carried out for each protocol. BM-MSCs from one dog were successfully cultured and expanded. Cells exposed to the two-step protocol appeared rarely grouped to form small clusters; gene expression analysis showed a slight increase in PDX-1 and insulin expression, but no insulin protein production nor secretion in the culture medium was detected either under basal conditions or following glucose stimulation. Conversely, cells exposed to the three-step protocol under a 3D culture system formed colony-like structures; insulin gene expression was upregulated compared to undifferentiated control and IPCs colonies secreted insulin in the culture medium, although insulin secretion was not enhanced by high-glucose culture conditions. The single experiment results suggest that the three-step differentiation protocol could generate IPCs from cBM-MSCs; however, further experiments are needed to confirm these data. The ability of IPCs from cBM- MSCs to produce insulin, described here for the first time, is a preliminary interesting result. Nevertheless, the IPCs' unresponsiveness to glucose, if confirmed, would affect its clinical application. Further studies are necessary to establish a differentiation protocol in this perspective.
在人类和兽医学中，使用产胰岛素细胞（IPC）的细胞疗法有望成为胰岛素注射或胰岛移植治疗糖尿病的替代治疗选择。已有多种将间充质干细胞（MSC）分化为IPC的方案被报道；迄今为止，仅从犬脂肪组织来源的MSC（c-MSC）中获得了葡萄糖反应性IPC，而犬骨髓来源的MSC（cBM-MSC）中尚未获得。因此，本研究旨在使用两种分化方案从cBM-MSC中体外生成葡萄糖反应性IPC：一种是使用曲古抑菌素（TSA）的两步方案，另一种是使用巯基乙醇诱导胰腺和十二指肠同源盒基因1（PDX-1）表达的三步方案。每个方案都进行了一次实验。成功培养并扩增了一只犬的BM-MSC。采用两步方案处理的细胞很少聚集成小簇；基因表达分析显示PDX-1和胰岛素表达略有增加，但在基础条件下或葡萄糖刺激后，培养基中均未检测到胰岛素蛋白的产生或分泌。相反，在三维培养系统中采用三步方案处理的细胞形成了集落样结构；与未分化对照相比，胰岛素基因表达上调，IPC集落在培养基中分泌胰岛素，尽管高糖培养条件并未增强胰岛素分泌。单次实验结果表明，三步分化方案可从cBM-MSC中生成IPC；然而，需要进一步实验来证实这些数据。首次在此描述的cBM-MSC来源的IPC产生胰岛素的能力是一个初步的有趣结果。然而，如果IPC对葡萄糖无反应得到证实，将影响其临床应用。从这个角度来看，有必要进行进一步研究以建立一种分化方案。
