Department of Surgery, Medicine, Dentistry and Morphological Sciences with Interest in Transplant, Oncology and Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy.
Dig Liver Dis. 2013 Aug;45(8):669-76. doi: 10.1016/j.dld.2013.02.007. Epub 2013 May 3.
To investigate the ability of human amniotic fluid stem cells and human dental pulp stem cells to differentiate into insulin-producing cells.
Human amniotic fluid stem cells and human dental pulp stem cells were induced to differentiate into pancreatic β-cells by a multistep protocol. Islet-like structures were assessed in differentiated human amniotic fluid stem cells and human dental pulp stem cells after 21 days of culture by dithizone staining. Pancreatic and duodenal homebox-1, insulin and Glut-2 expression were detected by immunofluorescence and confocal microscopy. Insulin secreted from differentiated cells was tested with SELDI-TOF MS and by enzyme-linked immunosorbent assay.
Human amniotic fluid stem cells and human dental pulp stem cells, after 7 days of differentiation started to form islet-like structures that became evident after 14 days of induction. SELDI-TOF MS analysis, revealed the presence of insulin in the media of differentiated cells at day 14, further confirmed by enzyme-linked immunosorbent assay after 7, 14 and 21 days. Both stem cell types expressed, after differentiation, pancreatic and duodenal homebox-1, insulin and Glut-2 and were positively stained by dithizone. Either the cytosol to nucleus translocation of pancreatic and duodenal homebox-1, either the expression of insulin, are regulated by glucose concentration changes. Day 21 islet-like structures derived from both human amniotic fluid stem cells and human dental pulp stem cell release insulin in a glucose-dependent manner.
The present study demonstrates the ability of human amniotic fluid stem cells and human dental pulp stem cell to differentiate into insulin-producing cells, offering a non-pancreatic, low-invasive source of cells for islet regeneration.
研究人羊水干细胞和人牙髓干细胞向胰岛素分泌细胞分化的能力。
通过多步诱导方案，将人羊水干细胞和人牙髓干细胞诱导分化为胰岛样细胞。培养 21 天后，通过二噻嗪染色评估分化后的人羊水干细胞和人牙髓干细胞中胰岛样结构的形成。通过免疫荧光和共聚焦显微镜检测胰腺和十二指肠同源盒-1、胰岛素和 Glut-2 的表达。通过 SELDI-TOF MS 和酶联免疫吸附试验检测分化细胞分泌的胰岛素。
人羊水干细胞和人牙髓干细胞在分化的第 7 天开始形成胰岛样结构，在诱导的第 14 天变得明显。SELDI-TOF MS 分析显示，在分化细胞的培养基中于第 14 天存在胰岛素，7、14 和 21 天后通过酶联免疫吸附试验进一步证实。两种干细胞在分化后均表达胰腺和十二指肠同源盒-1、胰岛素和 Glut-2，并被二噻嗪染色阳性染色。无论是胰腺和十二指肠同源盒-1 的细胞质到细胞核易位，还是胰岛素的表达，均受葡萄糖浓度变化的调节。第 21 天，人羊水干细胞和人牙髓干细胞来源的胰岛样结构以葡萄糖依赖的方式释放胰岛素。
本研究证明了人羊水干细胞和人牙髓干细胞向胰岛素分泌细胞分化的能力，为胰岛再生提供了一种非胰腺、低侵袭性的细胞来源。
