Single-injection ex ovo transplantation method for broad spinal cord engraftment of human pluripotent stem cell-derived motor neurons.
Single-injection ex ovo transplantation method for broad spinal cord engraftment of human pluripotent stem cell-derived motor neurons.
复制标题
单次注射卵外移植方法，用于人类多能干细胞衍生的运动神经元的广泛脊髓移植。
DOI：
10.1016/j.jneumeth.2018.01.006
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发表时间：
2018
影响因子：
3
通讯作者：
Ashton,RandolphS
中科院分区：
文献类型：
--
作者：
Estevez-Silva,MariaC;Sreeram,Akshitha;Cuskey,Stephanie;Fedorchak,Nikolai;Iyer,Nisha;Ashton,RandolphS
背景将人多能干细胞（human pluripotent stem cell，hPSC）来源的神经元移植到鸡胚中是评估其移植潜力的一种初步方法。然而，随着近年来在衍生不同的人类神经元亚型方面的进展，仍然需要优化和标准化针对特定亚型在其相关解剖部位的这种移植方法。并开发了一个单一的注射方案，植入hMN整个脊髓，使广泛的区域植入的可能性。注射到神经管腔中产生100%的鸡胚存活率和成功的移植率，从颈脊髓的喙侧到腰脊髓的尾侧观察到MN植入。与Pax 6 +/Nkx6.1+/Olig 2+祖细胞或成熟HB 9 +/ChAT + hMN相比，HB 9 +/ChAT − hMN前体的移植产生了最大量的植入。与现有方法的比较我们的单次注射hMN移植方法是第一个标准化鸡胚移植的最佳hMN表型，提供了植入定量的规则，结论将HB 9 +/ChAT−hMN前体细胞移植到Hamburger汉密尔顿（HH）期15- 20天的鸡胚中，18使用单次腔注射赋予胚胎存活和细胞移植在整个脊髓的不同区域中的高概率。
BackgroundTransplantation of human pluripotent stem cell (hPSC)-derived neurons into chick embryos is an established preliminary assay to evaluate engraftment potential. Yet, with recent advances in deriving diverse human neuronal subtypes, optimizing and standardizing such transplantation methodology for specific subtypes at their correlated anatomical sites is still required.New methodWe determined the optimal stage of hPSC-derived motor neuron (hMN) differentiation forex ovotransplantation, and developed a single injection protocol that implants hMNs throughout the spinal cord enabling broad regional engraftment possibilities.ResultsA single injection into the neural tube lumen yielded a 100% chick embryo survival and successful transplantation rate with MN engraftment observed from the rostral cervical through caudal lumbar spinal cord. Transplantation of HB9+/ChAT−hMN precursors yielded the greatest amount of engraftment compared to Pax6+/Nkx6.1+/Olig2+progenitors or mature HB9+/ChAT+hMNs.Comparison with existing method(s)Our single injection hMN transplant method is the first to standardize the optimal hMN phenotype for chick embryo transplantation, provide a rubric for engraftment quantification, and enable broad engraftment throughout the spinal cord with a single surgical intervention.ConclusionTransplantation of HB9+/ChAT−hMN precursors into chick embryos of Hamburger Hamilton (HH) stages 15–18 using a single luminal injection confers a high probability of embryo survival and cell engraftment in diverse regions throughout the spinal cord.