Tomita Koichi, Ishikawa Hiroshi
Department of Digestive and Transplantation Surgery, Tokyo Medical University Hachioji Medical Center, 1163 Tatemachi, Hachioji, Tokyo 193-0998, Japan.
Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
ScientificWorldJournal. 2019 Jan 9;2019:6264072. doi: 10.1155/2019/6264072. eCollection 2019.
Pluripotent stem cells are used in regenerative medicine and exist in various internal organs. However, there are a small number of reports of neural cells or neural stem cells existing in the spleen. In this study, we sought to identify possible neural stem cells in the mouse spleen. The spleens of ICR mice were removed and small specimens were incubated in Dulbecco's modified Eagle's medium with Nutrient Mixture F-12 containing either 10% fetal bovine serum (FBS), 20% FBS, 10% neonate bovine serum, or 10% fetal calf serum. Neural cell medium was also used. The cultured cells were investigated for expression of the neural cell markers neuron-specific enolase (NSE) and neurofilament 150 kDa (NF-150) by immunocytochemistry. Mouse spleens were also examined by immunohistochemistry for NSE, NF-150, NF-200, peripherin, and glial fibrillary acidic protein. Cells morphologically resembling neural cells were obtained and were positive for neural cell markers. Some of the cells generated sphere-like formations, which may have been neurospheres. Cell proliferation was best in medium containing 10% FBS. Cells positive for neural markers were observed in the subcapsular and perivascular regions of the spleen. The cells were round and present in much lower numbers than in cell culture. These cells are suspected neural stem cells and would be expected to differentiate into neural cells in cell culture. This report suggests the existence of neural stem cells in the mouse spleen.
多能干细胞被用于再生医学，存在于各种内脏器官中。然而，仅有少数关于脾脏中存在神经细胞或神经干细胞的报道。在本研究中，我们试图鉴定小鼠脾脏中可能存在的神经干细胞。切除ICR小鼠的脾脏，将小块标本置于含有10%胎牛血清（FBS）、20% FBS、10%新生牛血清或10%胎犊血清的杜氏改良 Eagle 培养基与营养混合物F-12中培养。也使用了神经细胞培养基。通过免疫细胞化学研究培养细胞中神经细胞标志物神经元特异性烯醇化酶（NSE）和150 kDa神经丝（NF-150）的表达。还通过免疫组织化学对小鼠脾脏进行NSE、NF-150、NF-200、外周蛋白和胶质纤维酸性蛋白检测。获得了形态上类似于神经细胞的细胞，且这些细胞对神经细胞标志物呈阳性反应。一些细胞形成了球形结构，可能是神经球。在含有10% FBS的培养基中细胞增殖最佳。在脾脏的被膜下和血管周围区域观察到了神经标志物呈阳性的细胞。这些细胞呈圆形，数量比细胞培养中的少得多。这些细胞被怀疑是神经干细胞，预计在细胞培养中会分化为神经细胞。本报告提示小鼠脾脏中存在神经干细胞。
