A novel, microfluidic high-throughput single-cell encapsulation of human bone marrow mesenchymal stromal cells

IF 4.2 3区 医学 Q2 ENGINEERING, BIOMEDICAL Journal of Materials Science: Materials in Medicine Pub Date : 2024-03-25 DOI:10.1007/s10856-024-06785-z
Narjes Rashidi, Alex Slater, Giordana Peregrino, Matteo Santin
{"title":"A novel, microfluidic high-throughput single-cell encapsulation of human bone marrow mesenchymal stromal cells","authors":"Narjes Rashidi,&nbsp;Alex Slater,&nbsp;Giordana Peregrino,&nbsp;Matteo Santin","doi":"10.1007/s10856-024-06785-z","DOIUrl":null,"url":null,"abstract":"<div><p>The efficacy of stem-cell therapy depends on the ability of the transplanted cells to escape early immunological reactions and to be retained at the site of transplantation. The use of tissue engineering scaffolds or injectable biomaterials as carriers has been proposed, but they still present limitations linked to a reliable manufacturing process, surgical practice and clinical outcomes. Alginate microbeads are potential candidates for the encapsulation of mesenchymal stromal cells with the aim of providing a delivery carrier suitable for minimally-invasive and scaffold-free transplantation, tissue-adhesive properties and protection from the immune response. However, the formation of stable microbeads relies on the cross-linking of alginate with divalent calcium ions at concentrations that are toxic for the cells, making control over the beads’ size and a single-cell encapsulation unreliable. The present work demonstrates the efficiency of an innovative, high throughput, and reproducible microfluidic system to produce single-cell, calcium-free alginate coatings of human mesenchymal stromal cells. Among the various conditions tested, visible light and confocal microscopy following staining of the cell nuclei by DAPI showed that the microfluidic system yielded an optimal single-cell encapsulation of 2000 cells/min in 2% w/v alginate microcapsules of reproducible morphology and an average size of 28.2 ± 3.7 µm. The adhesive properties of the alginate microcapsules, the viability of the encapsulated cells and their ability to escape the alginate microcapsule were demonstrated by the relatively rapid adherence of the beads onto tissue culture plastic and the cells’ ability to gradually disrupt the microcapsule shell after 24 h and proliferate. To mimic the early inflammatory response upon transplantation, the encapsulated cells were exposed to proliferating macrophages at different cell seeding densities for up to 2 days and the protection effect of the microcapsule on the cells assessed by time-lapse microscopy showing a shielding effect for up to 48 h. This work underscores the potential of microfluidic systems to precisely encapsulate cells by good manufacturing practice standards while favouring cell retention on substrates, viability and proliferation upon transplantation.</p><h3>Graphical Abstract</h3><div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":647,"journal":{"name":"Journal of Materials Science: Materials in Medicine","volume":"35 1","pages":""},"PeriodicalIF":4.2000,"publicationDate":"2024-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10963554/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Materials Science: Materials in Medicine","FirstCategoryId":"5","ListUrlMain":"https://link.springer.com/article/10.1007/s10856-024-06785-z","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"ENGINEERING, BIOMEDICAL","Score":null,"Total":0}
引用次数: 0

Abstract

The efficacy of stem-cell therapy depends on the ability of the transplanted cells to escape early immunological reactions and to be retained at the site of transplantation. The use of tissue engineering scaffolds or injectable biomaterials as carriers has been proposed, but they still present limitations linked to a reliable manufacturing process, surgical practice and clinical outcomes. Alginate microbeads are potential candidates for the encapsulation of mesenchymal stromal cells with the aim of providing a delivery carrier suitable for minimally-invasive and scaffold-free transplantation, tissue-adhesive properties and protection from the immune response. However, the formation of stable microbeads relies on the cross-linking of alginate with divalent calcium ions at concentrations that are toxic for the cells, making control over the beads’ size and a single-cell encapsulation unreliable. The present work demonstrates the efficiency of an innovative, high throughput, and reproducible microfluidic system to produce single-cell, calcium-free alginate coatings of human mesenchymal stromal cells. Among the various conditions tested, visible light and confocal microscopy following staining of the cell nuclei by DAPI showed that the microfluidic system yielded an optimal single-cell encapsulation of 2000 cells/min in 2% w/v alginate microcapsules of reproducible morphology and an average size of 28.2 ± 3.7 µm. The adhesive properties of the alginate microcapsules, the viability of the encapsulated cells and their ability to escape the alginate microcapsule were demonstrated by the relatively rapid adherence of the beads onto tissue culture plastic and the cells’ ability to gradually disrupt the microcapsule shell after 24 h and proliferate. To mimic the early inflammatory response upon transplantation, the encapsulated cells were exposed to proliferating macrophages at different cell seeding densities for up to 2 days and the protection effect of the microcapsule on the cells assessed by time-lapse microscopy showing a shielding effect for up to 48 h. This work underscores the potential of microfluidic systems to precisely encapsulate cells by good manufacturing practice standards while favouring cell retention on substrates, viability and proliferation upon transplantation.

Graphical Abstract

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
一种新型的微流体高通量单细胞人骨髓间充质基质细胞包被技术。
干细胞疗法的疗效取决于移植细胞能否躲过早期免疫反应并保留在移植部位。有人提出使用组织工程支架或可注射生物材料作为载体,但它们在可靠的制造工艺、手术实践和临床结果方面仍存在局限性。藻酸盐微珠是封装间充质基质细胞的潜在候选材料,其目的是提供一种适合微创和无支架移植、具有组织粘附性和免受免疫反应影响的输送载体。然而,稳定微珠的形成有赖于海藻酸盐与二价钙离子的交联,而这种交联的浓度对细胞来说是有毒的,因此对微珠大小的控制和单细胞包被并不可靠。本研究展示了一种创新、高通量、可重复的微流体系统在生产单细胞、无钙海藻酸盐包衣人类间充质基质细胞方面的效率。在测试的各种条件中,用 DAPI 对细胞核染色后的可见光和共聚焦显微镜显示,微流控系统能以 2000 个细胞/分钟的速度在 2% w/v 的藻酸盐微胶囊中产生最佳的单细胞包被效果,微胶囊的形态可重复,平均大小为 28.2 ± 3.7 µm。海藻酸盐微胶囊的粘附特性、封装细胞的存活能力以及细胞逃逸海藻酸盐微胶囊的能力通过珠子在组织培养塑料上相对较快的粘附以及细胞在 24 小时后逐渐破坏微胶囊外壳并增殖的能力得到了证明。为了模拟移植后的早期炎症反应,将封装的细胞暴露在不同细胞播种密度的增殖巨噬细胞中长达 2 天,并通过延时显微镜评估微胶囊对细胞的保护作用,结果表明微胶囊具有长达 48 小时的屏蔽作用。这项工作强调了微流体系统的潜力,即按照良好生产规范标准精确封装细胞,同时有利于细胞在基质上的保留、移植后的存活和增殖。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
Journal of Materials Science: Materials in Medicine
Journal of Materials Science: Materials in Medicine 工程技术-材料科学:生物材料
CiteScore
8.00
自引率
0.00%
发文量
73
审稿时长
3.5 months
期刊介绍: The Journal of Materials Science: Materials in Medicine publishes refereed papers providing significant progress in the application of biomaterials and tissue engineering constructs as medical or dental implants, prostheses and devices. Coverage spans a wide range of topics from basic science to clinical applications, around the theme of materials in medicine and dentistry. The central element is the development of synthetic and natural materials used in orthopaedic, maxillofacial, cardiovascular, neurological, ophthalmic and dental applications. Special biomedical topics include biomaterial synthesis and characterisation, biocompatibility studies, nanomedicine, tissue engineering constructs and cell substrates, regenerative medicine, computer modelling and other advanced experimental methodologies.
期刊最新文献
Performance evaluation of a low-cost Ti-Mo-Fe (TMF8) as a replacement for Ti-6Al-4V for internal fixation implants used in mandibular angular fractures: a finite element analysis study Biocompatibility and antibacterial properties of medical stainless steel and titanium modified by alumina and hafnia films prepared by atomic layer deposition Nano-titanium coating on glass surface to improve platelet-rich fibrin (PRF) quality Enhancing osteogenesis and mandibular defect repair with magnesium-modified acellular bovine bone matrix Shear bond strength between dental adhesive systems and an experimental niobium-based implant material
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1