Ramanathan Mrunalini, Rahman Md Mahbobur, Shijirbold Ankhtsetseg, Mahmod Md Rashel, Miyauchi Hiromi, Matsuzaki Yumi, Kanno Takahiro, Fujita Yuki
Department of Oral and Maxillofacial Surgery, Shimane University Faculty of Medicine, 89-1 Enya-cho, Izumo City 693-8501, Shimane, Japan.
Department of Anatomy and Developmental Biology, Shimane University Faculty of Medicine, 89-1 Enya-cho, Izumo City 693-8501, Shimane, Japan.
NeuroSci. 2025 May 2;6(2):39. doi: 10.3390/neurosci6020039.
Mesenchymal stem/stromal cells (MSCs) are non-hematopoietic, plastic-adherent, and self-renewing cells capable of in vitro trilineage differentiation into fat, bone, and cartilage tissue. Suggestively, MSCs have additional plasticity, as demonstrated by their ability to differentiate in vitro into myocytes, neuron-like cells, and hepatocytes. MSCs are ideal for therapeutic application owing to their numerous advantages; they exhibit limited growth and differentiation abilities, leading to heterogeneous cell populations with inconsistent functions. However, highly purified MSCs, namely, rapidly expanding clones (RECs) that are isolated by single-cell sorting, display uniform functionality. RECs have the potential to offer many benefits, such as transplantable cells for treating several disorders of bone, heart, peripheral nerves, brain, and other organs. This study aimed to assess the effects of RECs on the pheochromocytoma (PC12) cell line, a well-known neuronal cell model.
PC12 cells were cultured under the following conditions: co-culture with RECs, treatment with REC-derived conditioned medium (CM), or co-culture with RECs using Transwell inserts for 7 days. The cells were stained with anti-βIII-tubulin antibody; the lengths of neurites were measured by image analysis.
Regarding the co-culture with RECs, PC12's outgrowth was significantly increased. The RECs expressed nerve growth factor (NGF), a neurotrophic factor that could act on PC12 cells to trigger cellular differentiation.
Our findings suggest that RECs via direct culture, intercellular communication in Transwell culture, and RECs CM promoted PC12 cell survival and outgrowth via NGF signaling.
间充质干/基质细胞（MSCs）是非造血、贴壁生长且能自我更新的细胞，能够在体外向脂肪、骨和软骨组织进行三系分化。提示性地，MSCs具有额外的可塑性，这已通过它们在体外分化为肌细胞、神经元样细胞和肝细胞的能力得到证明。由于其众多优点，MSCs是治疗应用的理想选择；然而，它们的生长和分化能力有限，导致细胞群体功能不一致且异质性。然而，高度纯化的MSCs，即通过单细胞分选分离出的快速扩增克隆（RECs），具有统一的功能。RECs有潜力带来许多益处，例如可用于治疗多种骨、心脏、外周神经、脑和其他器官疾病的可移植细胞。本研究旨在评估RECs对嗜铬细胞瘤（PC12）细胞系（一种著名的神经元细胞模型）的影响。
PC12细胞在以下条件下培养：与RECs共培养、用RECs来源的条件培养基（CM）处理或使用Transwell小室与RECs共培养7天。细胞用抗βIII-微管蛋白抗体染色；通过图像分析测量神经突长度。
关于与RECs共培养，PC12的生长显著增加。RECs表达神经生长因子（NGF），这是一种神经营养因子，可作用于PC12细胞以触发细胞分化。
我们的研究结果表明，RECs通过直接培养、Transwell培养中的细胞间通讯以及RECs CM，通过NGF信号通路促进了PC12细胞的存活和生长。
