Cattaneo E, Magrassi L, Butti G, Santi L, Giavazzi A, Pezzotta S
Institute of Pharmacological Sciences, Universitá di Milano, Italy.
Brain Res Dev Brain Res. 1994 Dec 16;83(2):197-208. doi: 10.1016/0165-3806(94)00137-5.
Conditionally immortalized (temperature-sensitive) striatal-derived neuronal progenitor cell lines and primary neuroepithelial cells were transplanted into the CNS of gestational day 15-16 rat fetuses using an 'in utero' surgical procedure. Each fetus received 2.5-3 x 10(4) donor cells previously labelled in vitro by incubation with 5-bromo-2'-deoxyuridine (BrdU). At 5 days following transplantation, 69% of the fetuses were still alive. Engrafted cells were detected by BrdU immunohistochemistry, and the appearance of the engrafted cells and the time course of Nestin and PCNA expression were measured at 6, 24, 64 h and 5 days after transplantation. The evolution of Large T-Antigen immunoreactivity in engrafted temperature-sensitive (ts) cells was also evaluated at the above time intervals. The results indicate that the majority of the implanted cells were aggregated into clusters 24 h after transplantation. These clusters were not visible at 6 h, when most of the cells were isolated. The clusters were located in both the ventricles and parenchyma. These findings were common to both ts cells and striatal primary neuroepithelial cells. At 64 h and 5 days, isolated cells associated with the germinal layer and scattered throughout the parenchyma were also found. In the clusters, Nestin expression decreased proportionally with time following transplantation. Furthermore, Large T-Antigen immunoreactivity disappeared from ts cells between 6 and 24 h after transplantation. Finally, measurements of the temporal evolution of PCNA expression within the clusters indicate a progressive reduction in the mitotic activity of the transplanted cells. The results demonstrate that striatal primary neuroepithelial cells and conditionally immortalized neuronal progenitors can survive, migrate and/or compartimentalize into clusters whilst changing their antigenic properties and ability to proliferate.
利用“子宫内”外科手术，将条件永生化（温度敏感型）纹状体来源的神经元祖细胞系和原代神经上皮细胞移植到妊娠第15 - 16天的大鼠胎儿中枢神经系统中。每个胎儿接受2.5 - 3×10⁴个预先在体外与5 - 溴 - 2'-脱氧尿苷（BrdU）孵育标记的供体细胞。移植后5天，69%的胎儿仍存活。通过BrdU免疫组织化学检测植入细胞，并在移植后6、24、64小时和5天测量植入细胞的外观以及巢蛋白（Nestin）和增殖细胞核抗原（PCNA）表达的时间进程。在上述时间间隔也评估了植入的温度敏感型（ts）细胞中大T抗原免疫反应性的演变。结果表明，大多数植入细胞在移植后24小时聚集成团。这些细胞团在6小时时不可见，此时大多数细胞是分散的。细胞团位于脑室和实质内。这些发现对于ts细胞和纹状体原代神经上皮细胞都是共同的。在64小时和5天时，还发现了与生发层相关并散布在整个实质中的孤立细胞。在细胞团中，Nestin表达随移植后的时间成比例下降。此外，大T抗原免疫反应性在移植后6至24小时之间从ts细胞中消失。最后，对细胞团内PCNA表达时间演变的测量表明移植细胞的有丝分裂活性逐渐降低。结果表明，纹状体原代神经上皮细胞和条件永生化神经元祖细胞能够存活、迁移和/或聚集成团，同时改变它们的抗原特性和增殖能力。
