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目的探索脊髓内移植神经前体细胞的无创性活体示踪方法。方法培养人神经前体细胞系hNPC-TERT,采用逆转录聚合酶链反应(RT-PCR)、免疫荧光染色、放射性配基结合实验检测hNPC-TERT多巴胺D2受体在体内外的表达,移植于兔正常脊髓后,以D2受体拮抗剂“C- raclopride作为示踪剂,正电子发射计算机断层成像(PET)显像示踪兔活体脊髓和离体脊髓内移植的hNPC-TERT。HeLa细胞移植于脊髓作为对照组。结果体外培养和脊髓内移植第2天的hNPC- TERT表达D2受体。静脉注射11C-raclopride后PET显像,兔活体脊髓和离体脊髓内hNPC-TERT移植部位放射性积聚,对照组仅见本底影像。PET图像中,hNPC-TERT移植部位标准化摄取值明显高于Hela细胞移植部位(P<0．05)。离体脊髓放射性剖面曲线分析及脊髓切片免疫荧光染色进一步证实了PET显像结果。结论以11C-raclopride为示踪剂,PET显像可以示踪到脊髓内移植第2天的人神经前体细胞系HNPC-TERT。提供了一种以特异性表面标志为靶点进行PET显像的移植干细胞活体示踪方法。
Objective To explore the noninvasive methods of in vivo tracing of neural precursor cells in spinal cord. Methods Human neural progenitor cell line hNPC-TERT was cultured in vitro and in vivo. The expression of hNPC-TERT dopamine D2 receptor in vivo and in vitro was detected by reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence staining and radioactive ligand binding assay. After normal spinal cord of rabbit, hNPC-TERT.HeLa cells transplanted in living and isolated spinal cord of rabbits were labeled with positron emission tomography (PET) with D2 receptor antagonist “C-raclopride” as tracer In the spinal cord as a control group.Results The hNPC-TERT expressed D2 receptor in vitro and in spinal cord on day 2. PET imaging after intravenous injection of 11C-raclopride, radioactive accumulation of hNPC-TERT transplants in the living spinal cord and isolated spinal cord of rabbits , The control group only see the background images.PET images, hNPC-TERT transplanted site standardized uptake value was significantly higher than Hela cell transplants (P <0.05) .Extraction of spinal cord radioactivity profile curve analysis and spinal cord immunofluorescence staining further confirmed PET scans were performed.Conclusion PET imaging can track the human neural progenitor cell line HNPC-TERT on day 2 of spinal cord transplantation with 11C-raclopride as a tracer, Stem cell transplant tracer for PET imaging in vivo targeting.