Yokobori Shoji, Sasaki Kazuma, Kanaya Takahiro, Igarashi Yutaka, Nakae Ryuta, Onda Hidetaka, Masuno Tomohiko, Suda Satoshi, Sowa Kota, Nakajima Masataka, Spurlock Markus S, Onn Chieng Lee, Hazel Tom G, Johe Karl, Gajavelli Shyam, Fuse Akira, Bullock M Ross, Yokota Hiroyuki
Department of Emergency and Critical Care Medicine, Nippon Medical School, Tokyo, Japan.
Department of Neurological Science, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan.
Front Neurol. 2019 Feb 12;10:82. doi: 10.3389/fneur.2019.00082. eCollection 2019.
Human neural stem cells (hNSCs) transplantation in several brain injury models has established their therapeutic potential. However, the feasibility of hNSCs transplantation is still not clear for acute subdural hematoma (ASDH) brain injury that needs external decompression. Thus, the aim of this pilot study was to test feasibility using a rat ASDH decompression model with two clinically relevant transplantation methods. Two different methods, stereotactic injection and hNSC-embedded matrix seating on the brain surface, were attempted. Athymic rats were randomized to uninjured or ASDH groups (F344/NJcl-rnu/rnu, = 7-10/group). Animals in injury group were subjected to ASDH, and received decompressive craniectomy and 1-week after decompression surgery were transplanted with green fluorescent protein (GFP)-transduced hNSCs using one of two approaches. Histopathological examinations at 4 and 8 weeks showed that the GFP-positive hNSCs survived in injured brain tissue, extended neurite-like projections resembling neural dendrites. The transplantation group had greater engraftment of hNSCs than matrix embedding approach. Immunohistochemistry with doublecortin, NeuN, and GFAP at 8 weeks after transplantation showed that transplanted hNSCs remained as immature neurons and did not differentiate toward to glial cell lines. Motor function was assessed with rotarod, compared to control group ( = 10). The latency to fall from the rotarod in hNSC transplanted rats was significantly higher than in control rats (median, 113 s in hNSC vs. 69 s in control, = 0.02). This study first demonstrates the robust engraftment of transplanted hNSCs in a clinically-relevant ASDH decompression rat model. Further preclinical studies with longer study duration are warranted to verify the effectiveness of hNSC transplantation in amelioration of TBI induced deficits.
在多种脑损伤模型中，人神经干细胞（hNSCs）移植已证实了其治疗潜力。然而，对于需要进行外部减压的急性硬膜下血肿（ASDH）脑损伤，hNSCs移植的可行性仍不明确。因此，本初步研究的目的是使用大鼠ASDH减压模型和两种临床相关的移植方法来测试其可行性。尝试了两种不同的方法，即立体定向注射和将hNSCs嵌入基质置于脑表面。将无胸腺大鼠随机分为未受伤组或ASDH组（F344/NJcl-rnu/rnu，每组n = 7 - 10）。损伤组动物接受ASDH手术，并进行减压颅骨切除术，在减压手术后1周，使用两种方法之一将绿色荧光蛋白（GFP）转导的hNSCs进行移植。在4周和8周时进行组织病理学检查，结果显示GFP阳性的hNSCs在受损脑组织中存活，并伸出类似神经树突的神经突样突起。移植组的hNSCs植入情况比基质嵌入方法更好。移植后8周用双皮质素、NeuN和GFAP进行免疫组织化学检测显示，移植的hNSCs仍为未成熟神经元，未向神经胶质细胞系分化。与对照组（n = 10）相比，使用转棒试验评估运动功能。hNSCs移植大鼠从转棒上掉落的潜伏期显著高于对照大鼠（中位数，hNSCs组为113秒，对照组为69秒，P = 0.02）。本研究首次证明了在临床相关的ASDH减压大鼠模型中，移植的hNSCs能够稳固植入。有必要进行更长研究周期的进一步临床前研究，以验证hNSCs移植在改善创伤性脑损伤所致功能缺陷方面的有效性。
