Lu Hai-xia, Yang Zhi-qian, Jiao Qian, Wang Yuan-yuan, Wang Li, Yang Peng-bo, Chen Xin-lin, Zhang Peng-bo, Wang Peng, Chen Ming-xia, Lu Xiao-yun, Liu Yong
Neurol Res. 2014 Mar;36(3):207-14. doi: 10.1179/1743132813Y.0000000281.
Efforts have been made by tissue engineers to create a permissive environment for neural regeneration, and to enhance the efficiency of neural stem cell (NSC) transplantation. However, to acquire sufficient number of seed cells on the material appears to be the main obstacle to constructing functional transplantable NSC-biomaterial complexes. A culture system has been optimized in the current study to maintain the specific characteristics of NSCs/neural progenitor cells (NPCs) on the material and achieve sustaining increased multipotent seed cells.
The PHBHHx film was selected as biomaterial and the surface was firstly modified with NaOH treatment. The NSCs/NPCs isolated from the cerebral cortex of rat embryos were cultured on the treated PHBHHx films in growth medium containing 1%, 5%, and 10% fetal bovine serum (FBS). Then the attachment, survival, proliferation, and differentiation of NSCs/NPCs were assessed.
NaOH treatment significantly increased the hydrophilicity of PHBHHx and enhanced NSCs/NPCs attachment. On the treated PHBHHx film, NSCs/NPCs survived well and actively proliferated in the medium containing 1% FBS. After 7-14 days in culture, approximately two-thirds of cells remained as nestin and Sox2 positive NSCs/NPCs. However, in the medium containing 5% and 10% FBS, NSCs/NPCs proliferation was reduced and differentiation, particularly glial differentiation was significantly promoted.
Growth medium containing low concentration of FBS is favorable for maintaining the characteristics, in terms of self-renewal and multiple differentiation, of NSCs/NPCs on NaOH-treated PHBHHx films. This could be a useful method to construct functional transplantable NSCs/NPCs-biomaterial complex.
组织工程学领域的研究人员致力于为神经再生创造适宜的环境，并提高神经干细胞（NSC）移植的效率。然而，在材料上获得足够数量的种子细胞似乎是构建功能性可移植NSC-生物材料复合物的主要障碍。本研究优化了一种培养系统，以维持材料上NSC/神经祖细胞（NPC）的特定特性，并实现多能种子细胞数量的持续增加。
选用聚（3-羟基丁酸酯-co-3-羟基己酸酯）（PHBHHx）薄膜作为生物材料，首先用氢氧化钠处理对其表面进行改性。从大鼠胚胎大脑皮层分离得到的NSC/NPC在含有1%、5%和10%胎牛血清（FBS）的生长培养基中培养于经处理的PHBHHx薄膜上。然后评估NSC/NPC的附着、存活、增殖和分化情况。
氢氧化钠处理显著提高了PHBHHx的亲水性，增强了NSC/NPC的附着。在经处理的PHBHHx薄膜上，NSC/NPC在含有1% FBS的培养基中存活良好且能积极增殖。培养7-14天后，约三分之二的细胞仍为巢蛋白和Sox2阳性的NSC/NPC。然而，在含有5%和10% FBS的培养基中，NSC/NPC的增殖减少，分化，尤其是胶质细胞分化显著增强。
低浓度FBS的生长培养基有利于维持经氢氧化钠处理的PHBHHx薄膜上NSC/NPC的自我更新和多向分化特性。这可能是构建功能性可移植NSC/NPC-生物材料复合物的一种有用方法。
