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In vitro and in vivo Characterization of Tendon Stem Cells and Role of Stem Cells in Tendon Healing.

机译：肌腱干细胞的体外和体内表征以及干细胞在肌腱愈合中的作用。

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摘要

Tendon repair remains a great challenge due to current therapies cannot restore normal tendon function. Tendon-derived stem cells (TDSCs) have been isolated from tendon tissues and characterized in vitro in recent studies and provide new strategies for tendon repair. But what should we know about tendon stem cells before we use them to repair injured tendon?;Although stem cells that originate from different tissues share some common stem cell characteristics, they might also exhibit some tissue unique properties and hence functional differences. Therefore, we hypothesized that TDSCs have unique stemness properties compared with bone marrow-derived stem cells (BMSCs). There has been no study to compare the stemness properties of TDSCs and BMSCs. Clinical applications often require the in vitro expansion of stem cells. in vitro microenvironment also affects the stemness properties and therapeutic potential of stem cells. It is not clear if the stemness properties of TDSCs can be maintained and how long that they can be preserved during in vitro expansion. Moreover, successful stem cell-based repair therapies will require an understanding of tissue specific stem cells in vivo and their roles in the tissue repair. Tendon stem cells have not been described in details in vivo and it is unknown whether these endogenous stem cells participate in the tendon healing.;Therefore, in order to better make use of TDSCs for tendon repair, the objective of this study is to characterize the stemness properties of TDSCs compared with BMSCs and also to investigate the stemness limitation of TDSCs during culture in vitro for clinical use purpose. Furthermore, this study aims to identify the putative tendon stem cells in vivo and their role in tendon healing. This study would tell how much we should know about tendon stem cells in vitro and in vivo.;In the first part of the study, TDSCs and BMSCs were isolated from the same GFP Sprague-Dawley rat. TDSCs showed higher mensenchymal and pluripotent stem cell makers; clonogenicity; proliferative capacity; and tenogenic, osteogenic, chondrogenic, and adipogenic differentiation markers and multi-lineage differentiation potential than BMSCs. Compared with BMSCs, TDSCs shows great stemness properties and might be an alternative cell source for tendon regeneration.;In the second part of this study, the senescence-associated beta-galactosidase activity of TDSCs increased while their stem cell-related marker expression and the multi-lineage differentiation potential decreased during in vitro passaging. It suggests that researchers and clinicians need to consider the changes of stemness properties of TDSCs when multiplying them in vitro for tissue engineering.;In the third part of the study, IdU label-retaining method was used for the labeling of stem cells in vivo. We have identified quiescent stem cells as IdU label retaining cells (LRCs) at the peritenon, tendon mid-substance and tendon-bone junction. More LRCs were found at the peri-tenon and tendon-bone junction compared to the mid-substance. Some LRCs could be identified in the peri-vascular niche in the peri-tenon, suggesting that peri-vascular niche is one source of tendon stem cells. After injury, The LRC number and the expression of proliferative, tendon-related, pluripotency and pericyte-related markers in LRCs in the window wound increased, indicating that LRCs might be involved in tendon repair via cell migration, proliferation and differentiation.;In conclusion, our results have provided new findings about the understanding of tendon-derived stem cells including their stemness properties and their changes during the in vitro culture, as well as in vivo identity of tendon stem cells, which might facilitate the application of TDSCs in tissue engineering for tendon repair in the future.
机译：由于目前的疗法无法恢复正常的肌腱功能，肌腱修复仍然是一个巨大的挑战。肌腱来源的干细胞（TDSC）已从肌腱组织中分离出来，并在最近的研究中进行了体外表征，并为肌腱修复提供了新的策略。但是，在使用肌腱干细胞修复受损的肌腱之前，我们应该了解些什么？；尽管起源于不同组织的干细胞具有一些共同的干细胞特征，但它们也可能表现出某些组织独特的特性，因此功能也有所不同。因此，我们假设与骨髓来源的干细胞（BMSCs）相比，TDSCs具有独特的干性。尚无研究比较TDSC和BMSC的干性。临床应用通常需要体外扩增干细胞。体外微环境还影响干细胞的干性和治疗潜力。尚不明确TDSC的干性是否可以保持以及在体外扩增过程中可以保留多长时间。此外，成功的基于干细胞的修复疗法将需要了解体内组织特异性干细胞及其在组织修复中的作用。肌腱干细胞尚未在体内进行详细描述，尚不清楚这些内源性干细胞是否参与肌腱的愈合。因此，为了更好地利用TDSC修复肌腱，本研究的目的是表征TDSCs的干性与BMSCs的比较，并研究TDSC在体外培养过程中的干性局限性，以用于临床用途。此外，本研究旨在确定体内假定的肌腱干细胞及其在肌腱愈合中的作用。这项研究将告诉我们我们应该在体外和体内对肌腱干细胞有多少了解。 TDSC显示出更高的间充质和多能干细胞制造者。克隆性增殖能力；以及成肌，成骨，成软骨和成脂分化标志物以及多系分化潜能。与骨髓间充质干细胞相比，TDSCs具有强大的干性，可能是肌腱再生的另一种细胞来源。在本研究的第二部分中，TDSCs的衰老相关β-半乳糖苷酶活性增加，而它们的干细胞相关标志物表达和在体外传代过程中，多系分化潜能降低。这提示研究人员和临床医生在体外扩增TDSCs进行组织工程时，需要考虑其干性的变化。研究的第三部分，采用IdU标记保留方法对体内干细胞进行标记。我们已经将静止干细胞确定为腹膜，肌腱中段物质和肌腱-骨连接处的IdU标签保留细胞（LRC）。与中部相比，在腱周和腱-骨连接处发现了更多的LRC。在腱周的血管周围生态位中可以鉴定出一些LRC，这表明血管周围的生态位是肌腱干细胞的来源之一。损伤后，窗伤口LRC中的LRC数量和增殖，肌腱相关，多能性和周细胞相关标志物的表达增加，表明LRC可能通过细胞迁移，增殖和分化参与肌腱修复。 ，我们的结果为了解肌腱干细胞提供了新发现，包括其干性和体外培养过程中的变化，以及肌腱干细胞的体内特性，这可能有助于TDSC在组织工程中的应用用于将来的肌腱修复。

著录项

  • 作者

    Tan, Qi.;

  • 作者单位

    The Chinese University of Hong Kong (Hong Kong).;

  • 授予单位 The Chinese University of Hong Kong (Hong Kong).;
  • 学科 Medicine.;Biomedical engineering.;Kinesiology.
  • 学位 Ph.D.
  • 年度 2014
  • 页码 181 p.
  • 总页数 181
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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