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An Immortalized Rat Ventral Mesencephalic Cell Line RTC4 is Protective in a Rodent Model of Stroke

机译：永生的大鼠腹中脑细胞系RTC4在中风的啮齿动物模型中具有保护作用。

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One therapeutic approach to stroke is the transplantation of cells capable of trophic support, reinnervation and/or regeneration. Previously, we have described the use of novel truncated isoforms of SV40 large T antigen to generate unique cell lines from several primary rodent tissue types. Here we describe the generation of two cell lines, RTC3 and RTC4, derived from primary mesencephalic tissue using a fragment of mutant T antigen, T155c (cDNA) expressed from the RSV promoter. Both lines expressed the glial markers vimentin and S100β, but not the neuronal markers NeuN, MAP2 or β-III-tubulin. A screen for secreted trophic factors revealed substantially elevated levels of platelet-derived growth factor (PDGF) in RTC4, but not RTC3 cells. When transplanted into rat cortex, RTC4 cells survived for at least 22 days and expressed PDGF. Since PDGF has been reported to reduce ischemic injury, we examined the protective functions of RTC4 cells in an animal model of stroke. RTC4 or RTC3 cells, or vehicle, were injected into rat cortex 15–20 min prior to a 60-min middle cerebral artery ligation. Forty-eight hours later, animals were sacrificed and the stroke volume was assessed by triphenyl-tetrazolium chloride (TTC) staining. Compared to vehicle or RTC3 cells, transplanted RTC4 cells significantly reduced stroke volume. Overall, we generated a cell line with glial properties that produces PDGF and reduces ischemic injury in a rat model of stroke.
机译：一种治疗中风的方法是移植能够进行营养支持，重新神经支配和/或再生的细胞。以前，我们已经描述了使用SV40大T抗原的新型截短同工型从几种主要的啮齿动物组织类型中产生独特的细胞系。在这里，我们描述了使用突变T抗原T155c（cDNA）从RSV启动子表达的片段，从原代中脑组织衍生出两种细胞系RTC3和RTC4的产生。两种系均表达神经胶质标记波形蛋白和S100β，但不表达神经元标记NeuN，MAP2或β-III-微管蛋白。对分泌的营养因子的筛查显示，RTC4细胞（而非RTC3细胞）中血小板衍生的生长因子（PDGF）的水平显着升高。当移植到大鼠皮质中时，RTC4细胞存活至少22天并表达PDGF。由于已经报道了PDGF减少了缺血性损伤，因此我们在中风的动物模型中检查了RTC4细胞的保护功能。在大脑中动脉结扎60分钟之前的15至20分钟，将RTC4或RTC3细胞或媒介物注入大鼠皮质。 48小时后，处死动物并通过三苯基氯化四氮唑（TTC）染色评估中风量。与媒介物或RTC3细胞相比，移植的RTC4细胞显着减少了中风量。总的来说，我们在中风大鼠模型中产生了具有神经胶质特性的细胞系，该细胞系产生PDGF并减少缺血性损伤。

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