Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio, USA; School of Biomedical Sciences, Kent State University, Kent, Ohio, USA; Department of Biotechnology and Genetic Engineering, Philadelphia University, Amman, Jordan.
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio, USA; Department of Orthopaedic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio, USA; Instituto Universitario del Hospital Italiano de Buenos Aires, Buenos Aires, Argentina.
Cytotherapy. 2018 Mar;20(3):343-360. doi: 10.1016/j.jcyt.2017.11.013. Epub 2018 Feb 1.
Connective tissue progenitors (CTPs) embody the heterogeneous stem and progenitor cell populations present in native tissue. CTPs are essential to the formation and remodeling of connective tissue and represent key targets for tissue-engineering and cell-based therapies. To better understand and characterize CTPs, we aimed to compare the (i) concentration and prevalence, (ii) early in vitro biological behavior and (iii) expression of surface-markers and transcription factors among cells derived from marrow space (MS), trabecular surface (TS), and adipose tissues (AT).
Cancellous-bone and subcutaneous-adipose tissues were collected from 8 patients. Cells were isolated and cultured. Colony formation was assayed using Colonyze software based on ASTM standards. Cell concentration ([Cell]), CTP concentration ([CTP]) and CTP prevalence (P) were determined. Attributes of culture-expanded cells were compared based on (i) effective proliferation rate and (ii) expression of surface-markers CD73, CD90, CD105, SSEA-4, SSEA-3, SSEA-1/CD15, Cripto-1, E-Cadherin/CD324, Ep-CAM/CD326, CD146, hyaluronan and transcription factors Oct3/4, Sox-2 and Nanog using flow cytometry.
Mean [Cell], [CTP] and P were significantly different between MS and TS samples (P = 0.03, P = 0.008 and P= 0.0003), respectively. AT-derived cells generated the highest mean total cell yield at day 6 of culture-4-fold greater than TS and more than 40-fold greater than MS per million cells plated. TS colonies grew with higher mean density than MS colonies (290 ± 11 versus 150 ± 11 cell per mm; P = 0.0002). Expression of classical-mesenchymal stromal cell (MSC) markers was consistently recorded (>95%) from all tissue sources, whereas all the other markers were highly variable.
The prevalence and biological potential of CTPs are different between patients and tissue sources and lack variation in classical MSC markers. Other markers are more likely to discriminate differences between cell populations in biological performance. Understanding the underlying reasons for variation in the concentration, prevalence, marker expression and biological potential of CTPs between patients and source tissues and determining the means of managing this variation will contribute to the rational development of cell-based clinical diagnostics and targeted cell-based therapies.
结缔组织祖细胞（CTPs）体现了天然组织中存在的异质性干细胞和祖细胞群体。CTPs 对于结缔组织的形成和重塑至关重要，是组织工程和基于细胞的治疗的关键靶点。为了更好地理解和表征 CTPs，我们旨在比较（i）浓度和流行率，（ii）早期体外生物学行为和（iii）源自骨髓腔（MS）、小梁表面（TS）和脂肪组织（AT）的细胞的表面标志物和转录因子的表达。
从 8 名患者中收集松质骨和皮下脂肪组织。分离并培养细胞。使用基于 ASTM 标准的 Colonyze 软件测定集落形成。通过基于（i）有效增殖率和（ii）表达表面标志物 CD73、CD90、CD105、SSEA-4、SSEA-3、SSEA-1/CD15、Cripto-1、E-Cadherin/CD324、Ep-CAM/CD326、CD146、透明质酸和转录因子 Oct3/4、Sox-2 和 Nanog 的流式细胞术比较培养细胞的属性，来确定细胞浓度 ([Cell])、CTP 浓度 ([CTP]) 和 CTP 流行率 (P)。
MS 和 TS 样本之间的平均 [Cell]、[CTP] 和 P 差异均具有统计学意义（P=0.03、P=0.008 和 P=0.0003）。源自 AT 的细胞在培养的第 6 天产生的总细胞产量最高，比 TS 高 4 倍，比 MS 高 40 倍以上，每百万个细胞接种。TS 集落的生长密度高于 MS 集落（290±11 与 150±11 个细胞/毫米；P=0.0002）。所有组织来源的经典间充质基质细胞 (MSC) 标志物的表达均一致（>95%），而所有其他标志物的表达均高度可变。
CTPs 的流行率和生物学潜力在患者和组织来源之间存在差异，并且缺乏经典 MSC 标志物的变异性。其他标志物更有可能区分细胞群体在生物学性能方面的差异。了解患者和源组织中 CTPs 的浓度、流行率、标志物表达和生物学潜力变化的潜在原因，并确定管理这种变化的方法，将有助于基于细胞的临床诊断和靶向基于细胞的治疗的合理发展。
