Arora Aditya, Sriram M, Kothari Anjaney, Katti Dhirendra S
Department of Biological Sciences and Bioengineering, Indian Institute of Technology-Kanpur, Kanpur, Uttar Pradesh, India.
Department of Biological Sciences and Bioengineering, Indian Institute of Technology-Kanpur, Kanpur, Uttar Pradesh, India.
Cytotherapy. 2017 Jul;19(7):881-894. doi: 10.1016/j.jcyt.2017.04.003. Epub 2017 May 4.
Cell source plays a deterministic role in defining the outcome of a cell-based cartilage regenerative therapy and its clinical translational ability. Recent efforts in the direction of co-culture of two or more cell types attempt to combine the advantages of constituent cell types and negate their demerits.
We examined the potential of co-culture of infrapatellar fat pad-derived mesenchymal stromal cells (IFP MSCs) and articular chondrocytes (ACs) in plasma clots in terms of their ratios and culture formats for cartilage tissue engineering.
It was observed that IFP MSCs and ACs interact positively to produce a better quality hyaline cartilage-like matrix. While a supra-additive deposition of sulfated Glycosaminoglycans (sGAG), collagen type II, aggrecan and link protein was observed, deposition of collagen type I and X was sub-additive. (Immuno)-histologically similar cartilage was generated in vitro in IFP MSC:AC ratio of 50:50 and pure AC groups thus yielding a hyaline cartilage with 50% reduced requirement of ACs. Subsequently, we investigated if this response could be improved further by enabling better cell-cell interactions using scaffold-free systems such as self-assembled cartilage or by encapsulating cellular micro-aggregates in plasma clot. However, it was inferred that while self-assembly may have enabled better cell-cell interaction, poor cell survival negated its overall beneficial role, whereas the micro-aggregate group demonstrated highly heterogeneous matrix deposition within the construct, thus diminishing its translational utility. Overall, it was concluded that co-culture of IFP MSCs and ACs at a ratio of 50:50 within plasma clots demonstrated potential for cell-based cartilage regenerative therapy.
细胞来源在决定基于细胞的软骨再生治疗结果及其临床转化能力方面起着决定性作用。最近在两种或更多种细胞类型共培养方向上的努力试图结合组成细胞类型的优点并消除其缺点。
我们研究了髌下脂肪垫来源的间充质基质细胞(IFP MSCs)和关节软骨细胞(ACs)在血浆凝块中共培养的潜力,涉及它们在软骨组织工程中的比例和培养形式。
观察到IFP MSCs和ACs产生积极相互作用,以产生质量更好的透明软骨样基质。虽然观察到硫酸化糖胺聚糖(sGAG)、II型胶原蛋白、聚集蛋白聚糖和连接蛋白有超加性沉积,但I型和X型胶原蛋白的沉积是次加性的。在体外,IFP MSC:AC比例为50:50的组和纯AC组产生了(免疫)组织学上相似的软骨,从而使ACs的需求量减少了50%,产生了透明软骨。随后,我们研究了使用无支架系统(如自组装软骨)或通过将细胞微聚集体封装在血浆凝块中来实现更好的细胞间相互作用,是否可以进一步改善这种反应。然而,可以推断,虽然自组装可能实现了更好的细胞间相互作用,但细胞存活率低抵消了其总体有益作用,而微聚集体组在构建体内表现出高度异质性的基质沉积,从而降低了其转化效用。总体而言,得出的结论是,在血浆凝块中以50:50的比例共培养IFP MSCs和ACs显示出基于细胞的软骨再生治疗的潜力。