Rani Bhavna, Qian Hong, Johansson Staffan
Department of Medical Biochemistry and Microbiology (IMBIM), Biomedical Center, Uppsala University, P.O. Box 582, 751 23 Uppsala, Sweden.
Department of Medicine Huddinge, Center for Hematology and Regenerative Medicine, Karolinska Institute, Karolinska University Hospital, 141 86 Stockholm, Sweden.
Cells. 2025 Jun 19;14(12):932. doi: 10.3390/cells14120932.
Mesenchymal stem cells (MSCs) have a broad clinical potential, but their selection and expansion on plastic cause unknown purity and phenotypic alterations, reducing therapy efficiency. Furthermore, their behavior in non-adherent conditions during systemic transplantation remains poorly understood. The sphere formation from single cells is commonly used to assess stemness, but MSCs lack this ability, raising questions about their anchorage dependence for proliferation. We investigated whether bone marrow-derived MSCs can complete cytokinesis in non-adherent environments. Primary and bone marrow-derived MSCs were synchronized in early mitosis using nocodazole and were cultured on soft, rigid, or non-adherent surfaces. Both and MSCs displayed an ALIX (abscission licensor) recruitment to the midbody 40-90 min post-nocodazole release, regardless of the substrate adherence. Cells maintained for 4hr in the suspension remained viable, and daughter cells rapidly migrated apart upon the re-adhesion to fibronectin-coated surfaces, demonstrating cytokinesis completion in suspension. These findings distinguish MSCs from fibroblasts (which require adhesion for division), provide a more general stemness feature, and suggest that adhesion-independent cytokinesis is a trait relevant to the post-transplantation survival and tissue homing. This property may offer strategies to expand MSCs with an improved purity and functionality and to enhance engraftment by leveraging cell cycle manipulation to promote an early extracellular matrix deposition at target sites.
间充质干细胞（MSCs）具有广泛的临床应用潜力，但其在塑料培养皿上的筛选和扩增会导致纯度不明及表型改变，从而降低治疗效果。此外，在全身移植过程中，它们在非贴壁条件下的行为仍知之甚少。单细胞形成球体通常用于评估干性，但MSCs缺乏这种能力，这引发了人们对其增殖的锚定依赖性的质疑。我们研究了骨髓来源的MSCs是否能在非贴壁环境中完成胞质分裂。使用诺考达唑使原代和骨髓来源的MSCs在有丝分裂早期同步化，并将其培养在柔软、坚硬或非贴壁表面上。无论底物是否贴壁，在诺考达唑释放后40 - 90分钟，MSCs均显示ALIX（切割许可因子）募集到中体。在悬浮状态下维持4小时的细胞仍保持活力，并且子细胞在重新粘附到纤连蛋白包被的表面后迅速分开迁移，这表明在悬浮状态下完成了胞质分裂。这些发现将MSCs与成纤维细胞（成纤维细胞需要粘附才能分裂）区分开来，提供了更普遍的干性特征，并表明不依赖粘附的胞质分裂是与移植后存活和组织归巢相关的特性。这一特性可能为提高MSCs的纯度和功能来进行扩增，以及通过利用细胞周期调控促进靶位点早期细胞外基质沉积来增强植入提供策略。
