Background

Three-dimensional culture system of retinal explant is commonly used to study retinal ganglion cell (RGC) axon regeneration in vitro. The retinal explants fabricated by traditional procedure in culture system, however, are usually too small (merely 0.5 × 0.5 mm) to be easily detected or treated by current experimental techniques. Also, the constituents of culture medium have not been fully elucidated.

New method

A fabrication procedure was developed to enlarge the retinal explants and explore the reasonable concentration of fetal bovine serum (FBS) for evaluating axonal regeneration.

Results

There were no significant differences in the density or length of regenerative neurites in the retinal explants fabricated by traditional and modified procedures. Increased FBS concentrations promoted neurite regeneration, decreased RGCs apoptosis, and activated tyrosine kinase B (TrkB) receptors, all reaching a plateau at 1 % FBS.

Comparison with existing methods

Compared with traditional procedure, the modified fabrication procedure facilitates application of experimental techniques to retinal explants, increases the efficiency of obtaining observation area of regenerating neurites, and reduces the wastage of retinal tissues. The recommended FBS concentration determined in this study is shown to be more suitable for studying neuronal regeneration.

Conclusion

The retinal explants made by the modified fabrication procedure are successfully applied to the three-dimensional culture system, and presented several advantages over the traditional one. Furthermore, a preliminary experiment must be performed to determine the suitable concentration of FBS in each study to ensure accuracy and stability of the results obtained from the three-dimension retinal culture system.

中文翻译：

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在三维视网膜培养系统中改进的视网膜外植体制造程序和培养基的优化配方。

背景

视网膜外植体的三维培养系统通常用于体外研究视网膜神经节细胞（RGC）轴突再生。然而，在培养系统中通过传统程序制造的视网膜外植体通常太小（仅为0.5×0.5 mm），以致于无法通过当前的实验技术轻易地检测或处理。另外，还没有完全阐明培养基的成分。

新方法

开发了一种制造程序以扩大视网膜外植体并探索合理浓度的胎牛血清（FBS）以评估轴突再生。

结果

用传统方法和改良方法制造的视网膜外植体中，再生神经突的密度或长度没有显着差异。FBS浓度升高可促进神经突再生，减少RGC凋亡，并激活酪氨酸激酶B（TrkB）受体，在1％FBS时均达到平稳状态。

与现有方法的比较

与传统方法相比，改进的制造方法有利于将实验技术应用于视网膜外植体，提高了获得再生神经突的观察区域的效率，并减少了视网膜组织的浪费。在这项研究中确定的推荐FBS浓度显示更适合于研究神经元再生。

结论

改进后的制作方法制成的视网膜外植体已成功应用于三维培养系统，并具有优于传统培养系统的几个优点。此外，必须进行初步实验以确定每个研究中合适的FBS浓度，以确保从三维视网膜培养系统获得的结果的准确性和稳定性。