Gu Lihong, Liu Hongju, Wang Long, Fan Haokai, Zheng Xinli, Xu Tieshan, Jiang Qicheng, Zhou Teng, Shi Liuyong
Institute of Animal Science and Veterinary Medicine, Hainan Academy of Agricultural Sciences, Haikou, China.
School of Mechanical and Electrical Engineering, Hainan University, Haikou, China.
PLoS One. 2024 Dec 5;19(12):e0301309. doi: 10.1371/journal.pone.0301309. eCollection 2024.
Myoblasts are defined as stem cells containing skeletal muscle cell precursors. However, there are some challenges associated with the purification of myoblast samples, including long culture times and ease of bacterial contamination. In this study, we propose a microfluidic myoblast cell enrichment and purification platform based on the principle of deterministic lateral displacement (DLD). To achieve this, we designed a DLD chip with three outlets and tested it on 11-day-old (E11) Wenchang chicken pectoral muscle tissue. A cell suspension was prepared using the collagenase method, pretreated, and then passed into the designed DLD chip for myoblast enrichment and purification. In this study, the number of myoblasts and the diameter of myoblasts increased slowly before E9, and the diameter of myofibers decreased and the number of myofibers increased rapidly after E9. The period when the muscle fibers are most numerous is on the E12, and the period when the diameter of the muscle fibers begins to increase again after reaching its lowest point is also on the E12. After E12, the diameter of the muscle fibers increased and the number of muscle fibers decreased. At E12, myoblasts clustered and fused, and the proliferation of myoblasts was greatly reduced. E12 is both intact myoblasts and the most vigorous proliferation period, so the best time to determine isolation is E12. We attained a myoblast cell recovery rate of 80%, a target outlet collection purity of 99%, and a chip throughput of 50 μ m/min. In this paper, we innovate chips design according to specific geometries and functions for Wenchang chicken pectoral muscle tissue, so as to optimize the isolation and purification process of myoblasts. This study provides a novel and effective method for the isolation and purification of skeletal muscle myoblasts.
成肌细胞被定义为含有骨骼肌细胞前体的干细胞。然而,成肌细胞样本的纯化存在一些挑战,包括培养时间长和容易受到细菌污染。在本研究中,我们基于确定性侧向位移(DLD)原理提出了一种微流控成肌细胞富集和纯化平台。为此,我们设计了一种具有三个出口的DLD芯片,并在11日龄(E11)文昌鸡胸肌组织上进行了测试。使用胶原酶法制备细胞悬液,进行预处理,然后将其通入设计好的DLD芯片中进行成肌细胞的富集和纯化。在本研究中,E9之前成肌细胞数量和直径增长缓慢,E9之后肌纤维直径减小而数量迅速增加。肌纤维数量最多的时期是在E12,肌纤维直径达到最低点后再次开始增加的时期也是在E12。E12之后,肌纤维直径增加而数量减少。在E12时,成肌细胞聚集并融合,成肌细胞的增殖大大减少。E12既是完整成肌细胞存在又是增殖最旺盛的时期,所以确定分离的最佳时间是E12。我们获得了80%的成肌细胞回收率、99%的目标出口收集纯度以及50μm/min的芯片通量。在本文中,我们根据文昌鸡胸肌组织的特定几何形状和功能对芯片设计进行了创新,从而优化了成肌细胞的分离和纯化过程。本研究为骨骼肌成肌细胞的分离和纯化提供了一种新颖有效的方法。