Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA.
Department of Orthopedic Surgery, Cleveland Clinic, Cleveland, Ohio, USA.
Am J Sports Med. 2019 Dec;47(14):3521-3530. doi: 10.1177/0363546519880905. Epub 2019 Oct 31.
Cell-based therapy for cartilage repair is a promising approach and is becoming an established technique. Yet, there is no consensus on the optimal cell source.
To provide a donor-matched quantitative comparison of the connective tissue progenitors (CTPs) derived from cartilage (Outerbridge grade 1-3 [G1-2-3]), bone marrow aspirate concentrate (BMC), infrapatellar fat pad (IPFP), synovium, and periosteum with respect to (1) cell concentration ([Cell], cells/mL), (2) CTP prevalence (P, colonies per million cells), and (3) biological performance based on in vitro proliferation potential (cells per colony) colony density, and differentiation potential (expression of negatively charged extracellular matrix: glycosaminoglycan-rich extra cellular matrix [GAG-ECM]).
Descriptive laboratory study.
Tissues were obtained from 10 patients undergoing total knee arthroplasty (mean age, 59 years; women, n = 6). Automated quantitative colony-forming unit analysis was used to compare [Cell], P, and CTP biological performance across tissue sources.
[Cell] was highest in grade 3 cartilage ( = .002) and BMC ( = .001). Median P was highest in IPFP ( = .001), synovium ( = .003), and G1-2 cartilage ( = .02). Proliferation was highest in synovium-derived CTPs ( < .001). Median colony density was highest in G1-2-3 ( < .001). Median GAG-ECM was highest in G1-2-3 ( < .001). Within each patient, CTPs derived from all tissues were highly heterogeneous in biological performance as determined by cells per colony, density, and GAG-ECM.
Tissue sources differ in [Cell], P, and biological attributes. The data presented in this study suggest that cartilage (G1-2-3) is the preferred tissue source for cartilage repair based on P and GAG-ECM, followed by synovium, IPFP, BMC, and periosteum. However, due to the heterogeneous mixture of CTPs within each tissue source, there exists a subset of CTPs with biological performance similar to G1-2-3 cartilage, particularly in synovium and IPFP. Performance-based clonal selection and expansion of preferred CTPs and their progeny will potentially lead to improved cell population with predictive future.
Optimal tissue regeneration strategies will require informed decisions regarding which of the available tissue sources to use. Optimizing cell sourcing in any tissue may require separation of CTPs with preferred attributes from those with less desirable attributes. The heterogeneity manifest in the early stage of colony formation represents an opportunity for performance-based clone selection for clinical cell processing and manufacturing.
基于细胞的软骨修复疗法是一种很有前途的方法，并且正在成为一种成熟的技术。然而，对于最佳的细胞来源，尚无共识。
旨在提供源自软骨（Outerbridge 分级 1-3 [G1-2-3]）、骨髓抽吸物浓缩物（BMC）、髌下脂肪垫（IPFP）、滑膜和骨膜的结缔组织祖细胞（CTP）的供体匹配的定量比较，具体涉及（1）细胞浓度（[Cell]，细胞/mL），（2）CTP 流行率（P，每百万细胞的集落数），以及（3）基于体外增殖潜力的生物学性能（每个集落的细胞数）、集落密度和分化潜力（表达带负电荷的细胞外基质：糖胺聚糖丰富的细胞外基质[GAG-ECM]）。
描述性实验室研究。
从 10 名接受全膝关节置换术的患者中获取组织（平均年龄 59 岁；女性 n=6）。使用自动定量集落形成单位分析来比较组织来源的[Cell]、P 和 CTP 生物学性能。
[Cell]在 3 级软骨（=.002）和 BMC（=.001）中最高。IPFP（=.001）、滑膜（=.003）和 G1-2 软骨（=.02）的中位数 P 最高。滑膜来源的 CTPs 的增殖能力最高（<.001）。G1-2-3 的中位数集落密度最高（<.001）。G1-2-3 的中位数 GAG-ECM 最高（<.001）。在每个患者中，源自所有组织的 CTP 在生物学性能方面高度异质，如每个集落的细胞数、密度和 GAG-ECM 所示。
组织来源在[Cell]、P 和生物学特性方面存在差异。本研究中的数据表明，基于 P 和 GAG-ECM，软骨（G1-2-3）是软骨修复的首选组织来源，其次是滑膜、IPFP、BMC 和骨膜。然而，由于每个组织来源中 CTP 的混合不均匀，存在具有与 G1-2-3 软骨相似的生物学性能的 CTP 子集，特别是在滑膜和 IPFP 中。基于性能的克隆选择和优先 CTP 及其后代的扩增将有可能产生具有可预测未来的改良细胞群体。
优化的组织再生策略需要根据可用的组织来源做出明智的决策。在任何组织中优化细胞来源都可能需要将具有理想特性的 CTP 与具有较差特性的 CTP 分离。在集落形成的早期阶段表现出的异质性为基于性能的克隆选择提供了机会，可用于临床细胞处理和制造。
