Mukai Takeo, Mori Yuka, Shimazu Takahisa, Takahashi Atsuko, Tsunoda Hajime, Yamaguchi Satoru, Kiryu Shigeru, Tojo Arinobu, Nagamura-Inoue Tokiko
Division of Molecular of Therapy, Center for Advanced Medical Research, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan; Department of Cell Processing and Transfusion, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan.
Department of Cell Processing and Transfusion, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan.
Neuroscience. 2017 Jul 4;355:175-187. doi: 10.1016/j.neuroscience.2017.05.006. Epub 2017 May 11.
Intraventricular hemorrhage (IVH) is a frequent complication of preterm newborns, resulting in cerebral palsy and cognitive handicap as well as hypoxic ischemic encephalopathy and periventricular leukomalacia. In this study, we investigated the restorative effect on neonatal IVH by umbilical cord-derived mesenchymal stromal cells (UC-MSCs) cultured in serum-free medium (RM medium) for clinical application. UC-MSCs were cultured with αMEM medium supplemented with FBS or RM. A neonatal IVH mouse model at postnatal day 5 was generated by intraventricular injection of autologous blood, and mice were intravenously administered 1×10 UC-MSCs two days after IVH. Brain magnetic resonance imaging was performed at postnatal day 15, 22 and neurological behavioral measurements were performed at postnatal day 23, accompanied by histopathological analysis and cytokine bead assays in serum after IVH with or without UC-MSCs. Both UC-MSCs cultured with αMEM and RM met the criteria of MSCs and improved behavioral outcome of IVH mice. Moreover the RM group exhibited significant behavioral improvement compared to the control group. Histopathological analysis revealed UC-MSCs cultured with RM significantly attenuated periventricular reactive gliosis, hypomyelination, and periventricular cell death observed after IVH. Furthermore, human brain-derived neurotrophic factor and hepatocyte growth factor were elevated in the serum, cerebrospinal fluid and brain tissue of neonatal IVH model mice 24h after UC-MSCs administration. These results suggest UC-MSCs attenuate neonatal IVH by protecting gliosis and apoptosis of the injured brain, and intravenous injection of UC-MSCs cultured in RM may be feasible for neonatal IVH in clinic.
脑室内出血（IVH）是早产儿常见的并发症，可导致脑瘫、认知障碍以及缺氧缺血性脑病和脑室周围白质软化。在本研究中，我们调查了在无血清培养基（RM培养基）中培养的脐带间充质基质细胞（UC-MSCs）对新生儿IVH的修复作用，以用于临床应用。将UC-MSCs分别用添加胎牛血清（FBS）的αMEM培养基或RM培养基培养。通过脑室内注射自体血建立出生后第5天的新生儿IVH小鼠模型，IVH后两天给小鼠静脉注射1×10的UC-MSCs。在出生后第15天进行脑磁共振成像，在出生后第23天进行神经行为测量，并在IVH后有或无UC-MSCs的情况下，对血清进行组织病理学分析和细胞因子珠检测。用αMEM和RM培养的UC-MSCs均符合间充质干细胞的标准，并改善了IVH小鼠的行为结果。此外，与对照组相比，RM组的行为改善更为显著。组织病理学分析显示，用RM培养的UC-MSCs显著减轻了IVH后观察到的脑室周围反应性胶质增生、髓鞘形成减少和脑室周围细胞死亡。此外，在给予UC-MSCs后24小时，新生儿IVH模型小鼠的血清、脑脊液和脑组织中的人脑源性神经营养因子和肝细胞生长因子升高。这些结果表明，UC-MSCs通过保护受损脑的胶质增生和凋亡来减轻新生儿IVH，静脉注射在RM中培养的UC-MSCs在临床上可能对新生儿IVH可行。
