Critical Analytics for Manufacturing Personalized-Medicine Interdisciplinary Research Group, Singapore-MIT Alliance for Research and Technology, Singapore, Singapore.
Department of Orthopaedic Surgery, National University of Singapore, Singapore, Singapore.
Am J Sports Med. 2024 Feb;52(2):503-515. doi: 10.1177/03635465231214431. Epub 2024 Jan 8.
The functional heterogeneity of culture-expanded mesenchymal stem cells (MSCs) has hindered the clinical application of MSCs. Previous studies have shown that MSC subpopulations with superior chondrogenic capacity can be isolated using a spiral microfluidic device based on the principle of inertial cell focusing.
The delivery of microfluidic-enriched chondrogenic MSCs that are consistent in size and function will overcome the challenge of the functional heterogeneity of expanded MSCs and will significantly improve MSC-based cartilage repair.
Controlled laboratory study.
A next-generation, fully automated multidimensional double spiral microfluidic device was designed to provide more refined and efficient isolation of MSC subpopulations based on size. Analysis of in vitro chondrogenic potential and RNA sequencing was performed on size-sorted MSC subpopulations. In vivo cartilage repair efficacy was demonstrated in an osteochondral injury model in 12-week-old rats. Defects were implanted with MSC subpopulations (n = 6 per group) and compared with those implanted with unsegregated MSCs (n = 6). Osteochondral repair was assessed at 6 and 12 weeks after surgery by histological, micro-computed tomography, and mechanical analysis.
A chondrogenic MSC subpopulation was efficiently isolated using the multidimensional double spiral device. RNA sequencing revealed distinct transcriptomic profiles and identified differential gene expression between subpopulations. The delivery of a chondrogenic MSC subpopulation resulted in improved cartilage repair, as indicated by histological scoring, the compression modulus, and micro-computed tomography of the subchondral bone.
We have established a rapid, label-free, and reliable microfluidic protocol for more efficient size-based enrichment of a chondrogenic MSC subpopulation. Our proof-of-concept in vivo study demonstrates the enhanced cartilage repair efficacy of these enriched chondrogenic MSCs.
The delivery of microfluidic-enriched chondrogenic MSCs that are consistent in size and function can overcome the challenge of the functional heterogeneity of expanded MSCs, resulting in significant improvement in MSC-based cartilage repair. The availability of such rapid, label-free enriched chondrogenic MSCs can enable better cell therapy products for cartilage repair with improved treatment outcomes.
培养的间充质干细胞（MSCs）的功能异质性阻碍了 MSCs 的临床应用。先前的研究表明，基于惯性细胞聚焦原理的螺旋微流控装置可分离具有优越软骨形成能力的 MSC 亚群。
输送具有一致大小和功能的微流控富集软骨形成 MSC 将克服扩展 MSC 的功能异质性带来的挑战，并显著改善基于 MSC 的软骨修复。
对照实验室研究。
设计了一种下一代、全自动多维双螺旋微流控装置，以基于大小更精细、更有效地分离 MSC 亚群。对大小分选的 MSC 亚群进行体外软骨形成潜力分析和 RNA 测序。在 12 周龄大鼠的骨软骨损伤模型中证明了体内软骨修复效果。将 MSC 亚群（每组 6 个）植入缺陷中，并与未分离的 MSC （每组 6 个）进行比较。术后 6 周和 12 周通过组织学、微计算机断层扫描和力学分析评估骨软骨修复情况。
多维双螺旋装置高效分离出软骨形成 MSC 亚群。RNA 测序显示出不同的转录组谱，并确定了亚群之间的差异基因表达。输送软骨形成 MSC 亚群可改善软骨修复，表现为组织学评分、软骨下骨的压缩模量和微计算机断层扫描。
我们建立了一种快速、无标记、可靠的微流控方案，用于更高效地基于大小富集软骨形成 MSC 亚群。我们的体内概念验证研究表明，这些富集的软骨形成 MSC 具有增强的软骨修复效果。
输送具有一致大小和功能的微流控富集软骨形成 MSC 可以克服扩展 MSC 的功能异质性挑战，从而显著改善基于 MSC 的软骨修复。这种快速、无标记的富集软骨形成 MSC 的可用性可以为软骨修复提供更好的细胞治疗产品，从而改善治疗效果。
