Laboratory of Molecular Neurosurgery, Department of Stereotactic and Functional Neurosurgery, Neurocentre, Albert-Ludwigs-University of Freiburg, Freiburg, Germany.
Cell Transplant. 2011;20(10):1515-27. doi: 10.3727/096368910X547435. Epub 2010 Dec 22.
Embryonic striatal graft-mediated functional recovery in the rodent lesion model of Huntington's disease (HD) has been shown to correlate with the proportion of dopamine- and adenosine 3',5'-monophosphate-regulated phosphoprotein with a molecular weight of 32 kDa (DARPP-32)-positive neurons in the graft. The current study investigated the impact of graft distribution on the yield of DARPP-32-positive cells in the grafts following either single-tract or multitract cell delivery protocols using the microtransplantation approach. Cells derived from the whole ganglionic eminence of E15 rat embryos, ubiquitously expressing green fluorescent protein (GFP), were implanted into unilaterally QA-lesioned rat striatum either as 2 × 1.8 μl macrodeposits in a single tract, or as 18 × 0.2 μl microdeposits disseminated over six needle, multitract, penetrations. For both groups, an ultrathin glass capillary with an outer diameter of 50 μm was used. Histological assessment at 4 months after transplantation showed nearly twofold increase of DARRP-32-positive striatal-like neurons in the multitract compared to the single-tract group. However, the cellular make-up of the grafts did not translate into functional differences as tested in a basic spontaneous behavior test. Furthermore, the volumetric values for overall volume, DARPP-32-positive patches, and dopaminergic projection zones were similar between both groups. The results show that distribution of fetal striatal tissue in multiple submicroliter deposits provides for an increased yield of striatal-like neurons, potentially due to the enlargement of the graft-host border area intensifying the graft's exposure to host-derived factors. Furthermore, the use of embryonic tissue from GFP donors was validated in cell-based therapy studies in the HD model.
胚胎纹状体移植物介导的亨廷顿病（HD）啮齿动物病变模型中的功能恢复与移植物中多巴胺和腺苷 3'，5'-单磷酸调节的磷酸蛋白 32kDa（DARPP-32）阳性神经元的比例相关。目前的研究使用微移植方法，研究了在单通道或多通道细胞输送方案下，移植物分布对移植物中 DARPP-32 阳性细胞产量的影响。源自 E15 大鼠胚胎整个神经节隆起的细胞，普遍表达绿色荧光蛋白（GFP），被植入单侧 QA 损伤的大鼠纹状体中，作为单通道中的 2×1.8μl 大沉积，或作为 18×0.2μl 微沉积散布在六个针、多通道、穿透中。对于这两组，都使用了外径为 50μm 的超薄玻璃毛细管。移植后 4 个月的组织学评估显示，与单通道组相比，多通道组的 DARRP-32 阳性纹状体样神经元增加了近两倍。然而，作为基本自发行为测试进行的测试并未将移植物的细胞组成转化为功能差异。此外，两组之间的总体体积、DARPP-32 阳性斑块和多巴胺能投射区的体积值相似。结果表明，将胎儿纹状体组织分布在多个亚微升沉积中可增加纹状体样神经元的产量，这可能是由于移植物-宿主边界区域的扩大，增强了移植物对宿主来源因子的暴露。此外，在 HD 模型中的细胞治疗研究中，使用 GFP 供体的胚胎组织进行了验证。
