John Jeya Lisha, Singh Gurbind, Parasuraman Ganesh, Livingston Abel, Rebekah Grace, Daniel Alfred Job, Sathishkumar Solomon, Vinod Elizabeth
Department of Physiology, Christian Medical College, Vellore, Tamil Nadu, 632002, India.
Centre of Stem Cell Research, (a unit of BRIC-inStem, Bengaluru), Christian Medical College, Vellore, Tamil Nadu, India.
Histochem Cell Biol. 2025 Aug 20;163(1):82. doi: 10.1007/s00418-025-02416-y.
Chondroprogenitors derived from articular cartilage offer a promising approach for treating cartilage pathologies owing to their high chondrogenic and low hypertrophic potential. Optimizing holding conditions and parenteral solutions for transporting these cells from the processing to the transplantation site is crucial to enable their clinical application. This study assessed the viability, molecular phenotype maintenance, and differentiation potential of human fibronectin adhesion assay-derived chondroprogenitors (FAA-CPs) suspended in five parenteral solutions-(a) normal saline (NS), (b) plasma-lyte A, (c) 5% dextrose, (d) hyaluronic acid (HA), and (e) platelet-rich plasma (PRP) at 5 × 10 cells/ml and stored at 4 °C for 0, 6, and 12 h. FAA-CPs were isolated from nondiseased cartilage samples (n = 3). The assessments done included viability by Vi-CELL BLU assay and calcein AM-propidium iodide; surface chondrogenic marker expression; and differentiation potential by confirmatory staining. The cells exhibited positive mesenchymal stem cells (MSC) markers, moderate-to-high chondrogenic marker expression, and trilineage differentiation potential. Viability was preserved in NS, plasma-lyte A, 5% dextrose, and HA, but significantly declined in PRP. All groups retained multilineage potential, with higher Safranin-O uptake and collagen II accumulation in NS, plasma-lyte A, and 5% dextrose, suggesting enhanced chondrogenesis. Notably, 5% dextrose exhibited minimal collagen X accumulation, indicating low hypertrophic potential. NS, plasma-lyte A, and 5% dextrose poses to be optimal parenteral solutions for the formulation of chondroprogenitor suspensions, with a holding time of up to 12 h. Factoring lower hypertrophic potential, 5% dextrose seems to stand out among the other solutions as a cell-delivery vehicle for the treatment of cartilage diseases.
源自关节软骨的软骨祖细胞因其高软骨生成潜力和低肥大潜力，为治疗软骨病变提供了一种很有前景的方法。优化用于将这些细胞从处理部位运输到移植部位的保存条件和肠胃外溶液，对于实现其临床应用至关重要。本研究评估了悬浮于五种肠胃外溶液中的人纤连蛋白黏附试验衍生软骨祖细胞（FAA-CPs）的活力、分子表型维持和分化潜力，这五种溶液分别为：（a）生理盐水（NS）、（b）平衡液A、（c）5%葡萄糖、（d）透明质酸（HA）和（e）富血小板血浆（PRP），细胞浓度为5×10个细胞/毫升，并在4℃下保存0、6和12小时。FAA-CPs从无病变软骨样本（n = 3）中分离得到。所进行的评估包括通过Vi-CELL BLU分析和钙黄绿素AM-碘化丙啶检测活力；表面软骨生成标志物表达；以及通过确证染色检测分化潜力。这些细胞表现出阳性间充质干细胞（MSC）标志物、中度至高软骨生成标志物表达以及三系分化潜力。NS、平衡液A、5%葡萄糖和HA中细胞活力得以保留，但PRP中的细胞活力显著下降。所有组均保留了多系分化潜力，NS、平衡液A和5%葡萄糖组中番红O摄取量更高且胶原II积累更多，表明软骨生成增强。值得注意的是，5%葡萄糖组中胶原X积累最少，表明肥大潜力低。NS、平衡液A和5%葡萄糖似乎是用于配制软骨祖细胞悬液的最佳肠胃外溶液，保存时间长达12小时。考虑到较低的肥大潜力，5%葡萄糖在其他溶液中似乎脱颖而出，可作为治疗软骨疾病的细胞递送载体。
