Novel organotypic in vitro slice culture model for intraventricular hemorrhage of premature infants.
Novel organotypic in vitro slice culture model for intraventricular hemorrhage of premature infants.
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DOI：
10.1002/jnr.23102
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发表时间：
2012-11
影响因子：
4.2
通讯作者：
Ballabh, Praveen
中科院分区：
文献类型：
--
作者：
Vinukonda, Govindaiah;Hu, Furong;Upreti, Chirag;Ungvari, Zoltan;Zia, Muhammad T.;Stanton, Patric K.;Ballabh, Praveen
早产儿脑室内出血（IVH）的脑损伤机制尚不清楚，目前尚无预防早产儿脑损伤的治疗策略。因此，有必要开发一种体外器官型前脑切片培养模型，以推进这种疾病的机制研究和治疗发展。我们培养了E29幼兔前脑切片，并用中等量（50 μl）或大量（100 μl）自体血处理培养的切片（CS），以模拟中度和重度IVH。在培养开始后2周内，通过碘化丙啶染色、LDH水平、小胶质细胞密度、神经元变性、髓鞘形成和神经胶质增生来评价血液诱导的CS损伤。CS在体外（DIV）存活至少14天。血液的应用引起显著的神经细胞变性。与未处理的对照组相比，用50或100 μl血液处理的CS中变性细胞更丰富，LDH水平以剂量依赖性方式升高。在治疗后7天和14天，血液处理的CS中的小胶质细胞密度高于对照组。在血液处理的CS中，髓鞘形成减少，神经胶质增生增强。血液成分的选择性应用显示，与RBC处理的切片相比，用血浆处理的CS显示出更多的髓鞘形成不足和神经胶质增生。本研究开发并表征了一种新的IVH兔前脑切片培养模型，其表现出与IVH人类婴儿相似的神经病理学变化。重要的是，在IVH中，血浆似乎比红细胞诱导更大的白色物质损伤，表明血浆是神经毒性成分的来源。
Mechanisms of brain injury in intraventricular hemorrhage (IVH) of premature infants are elusive; and no therapeutic strategy exists to prevent the brain damage in these infants. Therefore, there is a need to develop an in vitro organotypic forebrain slice-culture model to advance mechanistic studies and therapeutic developments for this disorder. We cultured forebrain slices from E29 rabbit pups and treated the cultured slices (CS) with moderate (50 μl) or large (100 μl) amount of autologous blood to mimic moderate and severe IVH. Blood-induced damage to CS was evaluated for propidium-iodide staining, LDH levels, microglial density, neuronal degeneration, myelination, and gliosis over 2 weeks after the initiation of culture. CS were viable for at least 14 days in vitro (DIV). The application of blood induced significant neural cell degeneration. Degenerating cells were more abundant and LDH levels were elevated in a dose-dependent manner in CS treated with 50 or 100 μl of blood compared to untreated controls. Microglial density was higher in blood-treated CS compared to controls at both 7 and 14 days post-treatment. Myelination was reduced and gliosis enhanced in blood-treated CS. Selective application of blood fractions revealed that CS treated with plasma displayed more hypomyelination and gliosis compared to RBC-treated slices. This study developed and characterized a novel rabbit forebrain-slice culture model of IVH that exhibits neuropatholgical changes similar to human infants with IVH. Importantly, plasma appears to induce greater white matter damage than erythrocytes in IVH, indicating plasma a source of neurotoxic components.
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影响因子：
2.8
作者：
Niwa, Tetsu;de Vries, Linda S.;Benders, Manon J. N. L.;Takahara, Taro;Nikkels, Peter G. J.;Groenendaal, Floris
通讯作者：
Groenendaal, Floris
影响因子：
8.3
作者：
Verhagen, Elise A.;ter Horst, Hendrik J.;Bos, Arend F.
通讯作者：
Bos, Arend F.
影响因子：
8.8
作者：
Brambrink AM;Evers AS;Avidan MS;Farber NB;Smith DJ;Martin LD;Dissen GA;Creeley CE;Olney JW
通讯作者：
Olney JW
影响因子：
8.3
作者：
Georgiadis, Paraskevi;Xu, Hongmin;Ballabh, Praveen
通讯作者：
Ballabh, Praveen
影响因子：
6.1
作者：
Fujimoto, S;Katsuki, H;Akaike, A
通讯作者：
Akaike, A