Kassem Dina H, Kamal Mohamed M, El-Kholy Abd El-Latif G, El-Mesallamy Hala O
Biochemistry Department, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.
Gynecology and Obstetrics Department, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
Biochimie. 2016 Aug;127:187-95. doi: 10.1016/j.biochi.2016.05.019. Epub 2016 Jun 3.
Recently, there has been much attention towards generation of insulin producing cells (IPCs) from stem cells, especially from Wharton's jelly mesenchymal stem cells (WJ-MSCs). However, generation of mature IPCs remains a challenge. Assessment of generation of IPCs was usually done by examining β-cell markers, however, assessment of pluripotency/stem cell markers drew less attention. Therefore, the purpose of this study was to investigate the levels of pluripotency/stem cell markers during differentiation of WJ-MSCs into IPCs and the association of these levels with differentiation outcomes. WJ-MSCs were isolated, characterized then induced to differentiate into IPCs using three different protocols namely A, B and C. Differentiated IPCs were assessed by the expression of pluripotency/stem cell markers, together with β-cell markers using qRT-PCR, and functionally by measuring glucose stimulated insulin secretion. Differentiated cells from protocol A showed lowest expression of pluripotency/stem cell markers and relatively best GSIS. However, protocol B showed concomitant expression of pluripotency/stem cell and β-cell markers with relatively less insulin secretion as compared to protocol A. Protocol C failed to generate glucose-responsive IPCs. In conclusion, sustained expression of pluripotency/stem cell markers could be associated with the incomplete differentiation of WJ-MSCs into IPCs. A novel finding for which further investigations are warranted.
最近,从干细胞,尤其是从脐带来源的间充质干细胞(WJ-MSCs)中生成胰岛素分泌细胞(IPCs)受到了广泛关注。然而,生成成熟的IPCs仍然是一个挑战。对IPCs生成的评估通常是通过检测β细胞标志物来进行的,然而,对多能性/干细胞标志物的评估却较少受到关注。因此,本研究的目的是调查WJ-MSCs分化为IPCs过程中多能性/干细胞标志物的水平,以及这些水平与分化结果之间的关联。分离并鉴定WJ-MSCs,然后使用三种不同的方案,即方案A、方案B和方案C,诱导其分化为IPCs。通过qRT-PCR检测多能性/干细胞标志物以及β细胞标志物的表达来评估分化后的IPCs,并通过测量葡萄糖刺激的胰岛素分泌来进行功能评估。方案A分化的细胞显示多能性/干细胞标志物的表达最低,葡萄糖刺激的胰岛素分泌(GSIS)相对最佳。然而,方案B显示多能性/干细胞标志物和β细胞标志物同时表达,与方案A相比胰岛素分泌相对较少。方案C未能生成对葡萄糖有反应的IPCs。总之,多能性/干细胞标志物的持续表达可能与WJ-MSCs向IPCs的不完全分化有关。这是一个需要进一步研究的新发现。