检测内皮细胞相关的人源 DNA 可显示移植的人骨髓干细胞在 ALS 小鼠的 CNS 毛细血管中的嵌合。
Detection of endothelial cell-associated human DNA reveals transplanted human bone marrow stem cell engraftment into CNS capillaries of ALS mice.
机构信息
Center of Excellence for Aging and Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, United States; Department of Neurosurgery and Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, United States; Department of Molecular Pharmacology and Physiology, University of South Florida, Morsani College of Medicine, Tampa, FL, United States; Department of Pathology and Cell Biology, University of South Florida, Morsani College of Medicine, Tampa, FL, United States.
Center of Excellence for Aging and Brain Repair, University of South Florida, Morsani College of Medicine, Tampa, FL, United States.
出版信息
Brain Res Bull. 2021 May;170:22-28. doi: 10.1016/j.brainresbull.2021.01.020. Epub 2021 Feb 2.
Repairing the altered blood-CNS-barrier in amyotrophic lateral sclerosis (ALS) is imperative to prevent entry of detrimental blood-borne substances into the CNS. Cell transplantation with the goal of replacing damaged endothelial cells (ECs) may be a new therapeutic approach for barrier restoration. We showed positive effects of human bone marrow-derived CD34+ cells (hBM34+) and endothelial progenitor cells (hBM-EPCs) intravenous transplantation into symptomatic G93A SOD1 mutant mice on barrier reparative processes. These benefits mainly occurred by administered cells engraftment into vascular walls in ALS mice; however, additional studies are needed to confirm cell engraftment within capillaries. The aim of this investigation was to determine the presence of human DNA within microvascular ECs isolated from the CNS tissues of G93A SOD1 mutant mice treated with human bone marrow-derived stem cells. The CNS tissues were obtained from previously cell-treated and media-treated G93A mice at 17 weeks of age. Real-time PCR (RT-PCR) assay for detection of human DNA was performed in ECs isolated from mouse CNS tissue. Viability of these ECs was determined using the LIVE/DEAD viability/cytotoxicity assay. Results showed appropriate EC isolation as verified by immunoexpression of endothelial cell marker. Human DNA was detected in isolated ECs from cell-treated mice with greater concentrations in mice receiving hBM-EPCs vs. hBM34 cells. Also, higher numbers of live ECs were determined in mice treated with hBM-EPCs vs. hBM34 cells or media-injection. Results revealed that transplanted human cells engrafted into mouse capillary walls and efficaciously maintained endothelium function. These study results support our previous findings showing that intravenous administration of hBM-EPCs into symptomatic ALS mice was more beneficial than hBM34 cell treatment in repair of barrier integrity, likely due to replacement of damaged ECs in mouse CNS vessels. Based on this evidence, hBM-EPCs may be advanced as a cell-specific approach for ALS therapy through restored CNS barrier integrity.
修复肌萎缩侧索硬化症（ALS）中改变的血脑屏障对于防止有害的血源性物质进入中枢神经系统至关重要。细胞移植的目的是替代受损的内皮细胞（ECs），可能是一种恢复屏障的新治疗方法。我们已经证明，将人骨髓来源的 CD34+细胞（hBM34+）和内皮祖细胞（hBM-EPCs）静脉内移植到症状性 G93A SOD1 突变小鼠体内，对屏障修复过程有积极影响。这些益处主要通过给予的细胞在 ALS 小鼠的血管壁内移植而来；然而，需要进一步的研究来确认细胞在毛细血管内的移植。本研究的目的是确定用人类骨髓源性干细胞处理后的 G93A SOD1 突变小鼠的中枢神经系统组织中分离的微血管内皮细胞（EC）中是否存在人类 DNA。中枢神经系统组织取自先前用细胞和培养基处理过的 G93A 小鼠，在 17 周龄时获得。使用从小鼠中枢神经系统组织中分离的 EC 进行人 DNA 的实时 PCR（RT-PCR）检测。使用 LIVE/DEAD 活力/细胞毒性测定法确定这些 EC 的活力。结果表明，适当的 EC 分离得到了验证，因为内皮细胞标志物的免疫表达得到了证实。从细胞处理过的小鼠中分离的 EC 中检测到了人类 DNA，并且接受 hBM-EPC 治疗的小鼠中浓度高于接受 hBM34 细胞治疗的小鼠。此外，与 hBM34 细胞或培养基注射治疗的小鼠相比，接受 hBM-EPC 治疗的小鼠中的活 EC 数量更多。结果表明，移植的人类细胞在小鼠毛细血管壁中植入，并有效地维持了内皮功能。这些研究结果支持我们之前的发现，即静脉内给予 hBM-EPCs 到症状性 ALS 小鼠比 hBM34 细胞治疗更有利于修复屏障完整性，这可能是由于在小鼠中枢神经系统血管中替换了受损的 ECs。基于这一证据，hBM-EPCs 可以作为一种针对 ALS 治疗的细胞特异性方法得到推进，通过恢复中枢神经系统屏障的完整性。
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