基于声波叠加的反重力生物反应器水下三维干细胞球形悬浮培养。

IF 11.3 1区 医学 Q1 Medicine Biomaterials Research Pub Date : 2023-05-19 DOI:10.1186/s40824-023-00383-w
Jung Hwan Park, Ju-Ro Lee, Sungkwon Park, Yu-Jin Kim, Jeong-Kee Yoon, Hyun Su Park, Jiyu Hyun, Yoon Ki Joung, Tae Il Lee, Suk Ho Bhang
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引用次数: 1

摘要

背景:近年来,各种研究表明,在干细胞培养中,3D细胞球体比2D细胞具有许多优势。然而,传统的三维球体培养方法存在球体形成时间长、实验过程复杂等缺点和局限性。在这里,我们利用声悬浮作为细胞培养平台,克服了传统3D培养方法的局限性。方法:在反重力生物反应器中,连续驻声波形成压力场,用于人间充质干细胞(hMSCs)的三维培养。hMSCs在压力场中被捕获和聚集,形成球状体。采用电镜、免疫染色、聚合酶链反应、western blot等方法对反重力生物反应器中球体的结构、活力、基因和蛋白表达进行分析。将反重力生物反应器制备的造血干细胞球体注射到小鼠后肢缺血模型中。对残肢进行量化,评价hMSC球体的治疗效果。结果:反重力生物反应器中的声悬浮使球状体比常规悬滴法更快、更致密,导致造血干细胞血管内皮生长因子、血管生成素2等血管生成旁分泌因子上调。反重力生物反应器培养的hMSCs球体注射后对小鼠后肢缺血模型的治疗效果,包括保肢程度、毛细血管形成、纤维化和炎症的衰减,均优于常规吊滴法形成的球体。结论:我们的干细胞声悬浮培养系统将为未来的三维细胞培养系统提供一个新的平台。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Subaqueous 3D stem cell spheroid levitation culture using anti-gravity bioreactor based on sound wave superposition.

Background: Recently, various studies have revealed that 3D cell spheroids have several advantages over 2D cells in stem cell culture. However, conventional 3D spheroid culture methods have some disadvantages and limitations such as time required for spheroid formation and complexity of the experimental process. Here, we used acoustic levitation as cell culture platform to overcome the limitation of conventional 3D culture methods.

Methods: In our anti-gravity bioreactor, continuous standing sonic waves created pressure field for 3D culture of human mesenchymal stem cells (hMSCs). hMSCs were trapped and aggerated in pressure field and consequently formed spheroids. The structure, viability, gene and protein expression of spheroids formed in the anti-gravity bioreactor were analyzed by electron microscope, immunostaining, polymerase chain reaction, and western blot. We injected hMSC spheroids fabricated by anti-gravity bioreactor into the mouse hindlimb ischemia model. Limb salvage was quantified to evaluate therapeutic efficacy of hMSC spheroids.

Results: The acoustic levitation in anti-gravity bioreactor made spheroids faster and more compact compared to the conventional hanging drop method, which resulted in the upregulation of angiogenic paracrine factors of hMSCs, such as vascular endothelial growth factor and angiopoietin 2. Injected hMSCs spheroids cultured in the anti-gravity bioreactor exhibited improved therapeutic efficacy, including the degree of limb salvage, capillary formation, and attenuation of fibrosis and inflammation, for mouse hindlimb ischemia model compared to spheroids formed by the conventional hanging drop method.

Conclusion: Our stem cell culture system using acoustic levitation will be proposed as a new platform for the future 3D cell culture system.

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来源期刊
Biomaterials Research
Biomaterials Research Medicine-Medicine (miscellaneous)
CiteScore
10.20
自引率
3.50%
发文量
63
审稿时长
30 days
期刊介绍: Biomaterials Research, the official journal of the Korean Society for Biomaterials, is an open-access interdisciplinary publication that focuses on all aspects of biomaterials research. The journal covers a wide range of topics including novel biomaterials, advanced techniques for biomaterial synthesis and fabrication, and their application in biomedical fields. Specific areas of interest include functional biomaterials, drug and gene delivery systems, tissue engineering, nanomedicine, nano/micro-biotechnology, bio-imaging, regenerative medicine, medical devices, 3D printing, and stem cell research. By exploring these research areas, Biomaterials Research aims to provide valuable insights and promote advancements in the biomaterials field.
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