Song Kedong, Li Liying, Li Ruipeng, Lim Mayasari, Liu Peng, Liu Tianqing
State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian, 116024, China,
Appl Biochem Biotechnol. 2014 Jun;173(3):838-50. doi: 10.1007/s12010-014-0874-6. Epub 2014 Apr 12.
The Ca-alginate/gelatin (CAG) microbeads were prepared and evaluated through assays for their mechanical strength, permeability, and the feasibility as a cell carrier for in vitro culture of neural stem cells. The effects of different concentrations of sodium alginate, gelatin, and calcium chloride on the mechanical strength of CAG microbeads were determined using a self-made puncture force tester. Following this, the microbeads were immersed in DMEM media for a specified period to test its decay resistance. A diffusion model including a calculation formula of diffusion coefficient was built to investigate the diffusion of glucose and bovine serum albumin (BSA) through the wall of the microbeads. Furthermore, the feasibility of the microbeads for in vitro culture was identified using neural stem cells from Kunming mouse. Through a systematic approach and comprehensive analysis, the optimal gelatin conditions for microbead preparation were determined; the final combination of parameters of 1.5 % (wt%) sodium alginate (SA), 0.5 % (wt%) gelatin, and 4 % (wt%) CaCl2 were the best conditions for NSC cultures. This experiment demonstrated that CAG microbeads had good cytocompatibility that made it suitable for the culture and successfully maintained stemness of neural stem cells.
制备了海藻酸钙/明胶（CAG）微珠，并通过检测其机械强度、渗透性以及作为神经干细胞体外培养细胞载体的可行性进行评估。使用自制的穿刺力测试仪测定不同浓度的海藻酸钠、明胶和氯化钙对CAG微珠机械强度的影响。在此之后，将微珠浸入DMEM培养基中一段特定时间以测试其抗降解性。建立了一个包括扩散系数计算公式的扩散模型，以研究葡萄糖和牛血清白蛋白（BSA）通过微珠壁的扩散。此外，使用来自昆明小鼠的神经干细胞确定了微珠用于体外培养的可行性。通过系统的方法和综合分析，确定了微珠制备的最佳明胶条件；1.5%（重量百分比）海藻酸钠（SA）、0.5%（重量百分比）明胶和4%（重量百分比）氯化钙的最终参数组合是神经干细胞培养的最佳条件。该实验表明，CAG微珠具有良好的细胞相容性，使其适合培养并成功维持神经干细胞的干性。
