Application of Fibronectin-Coated Microparticles for Isolation of Human iPSC-Derived MSCs.

IF 5.1 4区 医学 Q2 CELL & TISSUE ENGINEERING
Jae Hyeok Jang, In Sun Hwang, Ji Seob Kim, Sunjun Lee, Min Hee Moon, Bowon Kim, Seo Jeong Kim, Jongho Choi, Young Her, Soo-Hong Lee, Byung-Hyun Cha
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引用次数: 0

Abstract

Background: Bone marrow-derived MSCs (BMMSCs) are limited by availability, donor variability, and age-related decline, highlighting the need for alternative MSC sources. Induced pluripotent stem cells (iPSCs) offer a scalable solution. This study introduces a novel three-dimensional culture platform based on fibronectin (FN)-coated microparticles to efficiently derive and expand human iPSC-derived MSCs (iMSCs).

Methods: The system utilizes FN-coated non-porous (CytoDex) and porous (CytoPore) microparticles. FN coating efficacy on microparticle was confirmed using Rhodamine-labeled FN and confocal microscope. Base on various molecular cell biological experiment, the utilization of FN-coated CytoDex and CytoPore leverages an expanded surface area to improve iMSCs isolation. The characteristics and cell behaviors of iMSCs generated from FN-coated CytoDex and CytoPore was evaluated by immunophenotype analysis, cell proliferation, and senescence related assays. In vivo tissue regeneration was evaluated by microcomputed tomography and histopathological analyses.

Results: The porous microparticle CytoPore significantly enhanced FN coating efficiency, cell attachment, and proliferation compared to CytoDex. FN-coated CytoPore enabled the selective isolation of a high-purity MSC population from spontaneously differentiated iPSCs (SD-iPSCs) by day 7, confirmed by distinct morphology and mesodermal marker expression. The resulting iMSCs exhibited immunophenotypic characteristics comparable to adult MSCs, along with superior proliferative capacity, extended telomere length, and minimal senescence over 10 passages, in contrast to BMMSCs. Furthermore, iMSCs demonstrated effective in vivo tissue regeneration in an osteochondral defect model.

Conclusion: This novel FN-coated microparticle-based 3D culture platform enables efficient, large-scale production of high-quality iMSCs and holds strong potential for clinical applications in stem cell therapeutics and regenerative medicine.

纤维连接蛋白包被微颗粒在分离人间充质干细胞中的应用。
背景:骨髓来源的间充质干细胞(BMMSCs)受到可用性、供体可变性和年龄相关衰退的限制,突出了对替代间充质干细胞来源的需求。诱导多能干细胞(iPSCs)提供了一个可扩展的解决方案。本研究介绍了一种基于纤维连接蛋白(FN)包被微颗粒的新型三维培养平台,以有效地衍生和扩增人ipsc衍生的MSCs (iMSCs)。方法:该系统采用fn包被的无孔(CytoDex)和多孔(CytoPore)微粒。采用罗丹明标记FN和共聚焦显微镜验证FN对微粒的涂膜效果。在各种分子细胞生物学实验的基础上,利用fn包被的CytoDex和CytoPore利用扩大的表面积来提高iMSCs的分离。通过免疫表型分析、细胞增殖和衰老相关试验评估fn包被的CytoDex和CytoPore生成的iMSCs的特性和细胞行为。通过显微计算机断层扫描和组织病理学分析评估体内组织再生。结果:与CytoDex相比,多孔微粒CytoPore显著提高了FN包覆效率、细胞附着和增殖能力。fn包被的CytoPore能够在第7天从自发分化的iPSCs (SD-iPSCs)中选择性分离出高纯度的MSC群体,通过不同的形态和中胚层标记物表达证实了这一点。与BMMSCs相比,由此产生的iMSCs具有与成人MSCs相当的免疫表型特征,以及优越的增殖能力、延长的端粒长度和最小的衰老。此外,在骨软骨缺损模型中,iMSCs显示出有效的体内组织再生。结论:这种新型的fn包被微颗粒三维培养平台能够高效、大规模地生产高质量的iMSCs,在干细胞治疗和再生医学的临床应用中具有巨大的潜力。
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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来源期刊
Tissue engineering and regenerative medicine
Tissue engineering and regenerative medicine CELL & TISSUE ENGINEERING-ENGINEERING, BIOMEDICAL
CiteScore
6.80
自引率
5.60%
发文量
83
审稿时长
6-12 weeks
期刊介绍: Tissue Engineering and Regenerative Medicine (Tissue Eng Regen Med, TERM), the official journal of the Korean Tissue Engineering and Regenerative Medicine Society, is a publication dedicated to providing research- based solutions to issues related to human diseases. This journal publishes articles that report substantial information and original findings on tissue engineering, medical biomaterials, cells therapy, stem cell biology and regenerative medicine.
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