三维重建骨髓基质培养通过 STAT3 依赖性机制提高原代骨髓瘤细胞的体外活力。

Three-Dimensional Reconstructed Bone Marrow Matrix Culture Improves the Viability of Primary Myeloma Cells In-Vitro via a STAT3-Dependent Mechanism.

机构信息

Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB T6G 2E1, Canada.

Electron Microscopy Center, Basic Medical Science College, Harbin Medical University, Harbin 150080, China.

出版信息

Curr Issues Mol Biol. 2021 Jun 8;43(1):313-323. doi: 10.3390/cimb43010026.

DOI:10.3390/cimb43010026

PMID:34201211

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8928965/

Abstract

Primary myeloma (PM) cells are short-lived in conventional culture, which limited their usefulness as a study model. Here, we evaluated if three-dimensional (3D) culture can significantly prolong the longevity of PM cells in-vitro. We employed a previously established 3D model for culture of bone marrow mononuclear cells isolated from 15 patients. We assessed the proportion of PM cells, viability and proliferation using CD38 staining, trypan blue exclusion assays and carboxy fluorescein succinimidyl ester (CFSE) staining, respectively. We observed significantly more CD38+ viable cells in 3D than in conventional culture (65% vs. 25%, = 0.006) on day 3. CFSE staining showed no significant difference in cell proliferation between the two culture systems. Moreover, we found that PM cells in 3D culture are more STAT3 active by measure of pSTAT3 staining (66% vs. 10%, = 0.008). Treatment of IL6, a STAT3 activator significantly increased CD38+ cell viability (41% to 68%, = 0.021). In comparison, inhibition of STAT3 with Stattic significantly decreased PM cell viability in 3D culture (38% to 17% = 0.010). Neither IL6 nor Stattic affected the PM cell viability in conventional culture. This study suggests that 3D culture can significantly improve the longevity of PM cells in-vitro, and STAT3 activation can further improve their viability.

摘要

原发性骨髓瘤（PM）细胞在常规培养中寿命较短，这限制了它们作为研究模型的用途。在这里，我们评估了三维（3D）培养是否可以显著延长 PM 细胞在体外的寿命。我们采用了先前建立的从 15 名患者分离的骨髓单核细胞 3D 培养模型。我们分别使用 CD38 染色、台盼蓝排斥试验和羧基荧光素琥珀酰亚胺酯（CFSE）染色评估 PM 细胞的比例、活力和增殖。我们发现，在第 3 天，3D 培养中的 CD38+活细胞比例明显高于常规培养（65%比 25%， = 0.006）。CFSE 染色显示两种培养系统的细胞增殖无显著差异。此外，我们发现 3D 培养中的 PM 细胞通过 pSTAT3 染色测量具有更高的 STAT3 活性（66%比 10%， = 0.008）。用 STAT3 激活剂 IL6 处理可显著增加 CD38+细胞活力（从 41%增加到 68%， = 0.021）。相比之下，用 Stattic 抑制 3D 培养中的 STAT3 会显著降低 PM 细胞的活力（从 38%降至 17%， = 0.010）。IL6 或 Stattic 均未影响常规培养中的 PM 细胞活力。本研究表明，3D 培养可显著提高 PM 细胞在体外的寿命，而 STAT3 激活可进一步提高其活力。