Teshima Takahiro, Okamoto Keiji, Dairaku Kazuho, Nagashima Tomokazu, Michishita Masaki, Suzuki Ryohei, Matsumoto Hirotaka, Koyama Hidekazu
Laboratory of Veterinary Internal Medicine, Department of Veterinary Clinical Medicine, School of Veterinary Medicine, Faculty of Veterinary Science, Nippon Veterinary and Life Science University, 1-7-1 Kyonan-cho, Musashino, Tokyo 180-8602, Japan.
Research Center for Animal Life Science, Nippon Veterinary and Life Science University, 1-7-1 Kyonan-cho, Musashino, Tokyo 180-8602, Japan.
Stem Cells Int. 2020 Oct 18;2020:8841865. doi: 10.1155/2020/8841865. eCollection 2020.
The potential of mesenchymal stem cells (MSCs) to differentiate into nonmesodermal cells such as pancreatic beta cells has been reported. New cell-based therapy using MSCs for diabetes mellitus is anticipated as an alternative treatment option to insulin injection or islet transplantation in both human and veterinary medicine. Several protocols were reported for differentiation of MSCs into insulin-producing cells (IPCs), but no studies have reported IPCs generated from canine MSCs. The purpose of this study was to generate IPCs from canine adipose tissue-derived MSCs (AT-MSCs) and to investigate the effects of IPC transplantation on diabetic mice . Culturing AT-MSCs with the differentiation protocol under a two-dimensional culture system did not produce IPCs. However, spheroid-like small clusters consisting of canine AT-MSCs and human recombinant peptide -pieces developed under a three-dimensional (3D) culture system were successfully differentiated into IPCs. The generated IPCs under 3D culture condition were stained with dithizone and anti-insulin antibody. Canine IPCs also showed gene expression typical for pancreatic beta cells and increased insulin secretion in response to glucose stimulation. The blood glucose levels in streptozotocin-induced diabetic mice were decreased after injection with the supernatant of canine IPCs, but the hyperglycemic states of diabetic mice were not improved after transplanting IPCs subcutaneously or intramesenterically. The histological examination showed that the transplanted small clusters of IPCs were successfully engrafted to the mice and included cells positive for insulin by immunofluorescence. Several factors, such as the transplanted cell number, the origin of AT-MSCs, and the differentiation protocol, were considered potential reasons for the inability to improve the hyperglycemic state after IPC transplantation. These findings suggest that canine AT-MSCs can be differentiated into IPCs under a 3D culture system and IPC transplantation may be a new treatment option for dogs with diabetes mellitus.
已有报道称间充质干细胞（MSCs）具有分化为非中胚层细胞（如胰腺β细胞）的潜力。在人类和兽医学中，使用MSCs进行糖尿病新的细胞疗法有望成为胰岛素注射或胰岛移植的替代治疗选择。已有多种将MSCs分化为胰岛素产生细胞（IPCs）的方案被报道，但尚无研究报道从犬MSCs生成IPCs。本研究的目的是从犬脂肪组织来源的MSCs（AT-MSCs）生成IPCs，并研究IPCs移植对糖尿病小鼠的影响。在二维培养系统下按照分化方案培养AT-MSCs未能产生IPCs。然而，在三维（3D）培养系统下由犬AT-MSCs和人重组肽片段形成的类球体小簇成功分化为IPCs。在3D培养条件下生成的IPCs用双硫腙和抗胰岛素抗体染色。犬IPCs也显示出胰腺β细胞典型的基因表达，并在葡萄糖刺激下胰岛素分泌增加。注射犬IPCs的上清液后，链脲佐菌素诱导的糖尿病小鼠的血糖水平降低，但皮下或肠系膜内移植IPCs后糖尿病小鼠的高血糖状态并未改善。组织学检查显示，移植的IPCs小簇成功植入小鼠体内，免疫荧光显示其中包含胰岛素阳性细胞。移植细胞数量、AT-MSCs来源和分化方案等多种因素被认为是IPCs移植后无法改善高血糖状态的潜在原因。这些发现表明，犬AT-MSCs在3D培养系统下可分化为IPCs，IPCs移植可能是糖尿病犬的一种新的治疗选择。
