From the Cancer Cell Biology Group, Institut d'Investigació Sanitària Illes Balears; Servei de Genètica, Hospital Universitari Son Espases; Institut Català d'Oncologia, Hospital Germans Trias i Pujol; Cell Pro Tech Spain; and the University of Florida College of Medicine.
Plast Reconstr Surg. 2020 Dec;146(6):1285-1293. doi: 10.1097/PRS.0000000000007342.
Regenerative cell strategies rely on stromal cell implants to attain an observable clinical outcome. However, the effective cell dose to ensure a therapeutic response remains unknown. To achieve a higher cell dose, the authors hypothesized that reducing the volume occupied by mature adipocytes in lipoaspirate will concentrate the stromal vascular fraction present in the original tissue.
Human standardized lipoaspirate (n = 6) was centrifuged (1200 g for 3 minutes) and the water phase was discarded. Mechanical disaggregation was achieved by shearing tissue through 2.4- and 1.2-mm Luer-to-Luer transfers. After a second centrifugation (800 g for 10 minutes), stromal cell aggregates were separated from the supernatant oil phase. Lipoaspirate percentage composition was determined by its constituent weights. Cell content was measured by total DNA quantification, and partial cell viability was determined by image cytometry. Tissue sections were evaluated histologically (hematoxylin and eosin and Masson trichrome stains).
Stromal cell aggregates reduced the standardized lipoaspirate mass to 28.6 ± 4.2 percent. Accordingly, the cell density increased by 222.6 ± 63.3 percent (from 9.9 ± 1.4 million cells/g to 31.3 ± 6.6 million cells/g; p < 0.05). Cell viability was unaffected in stromal cell aggregates (71.3 ± 2.5 percent) compared to standardized lipoaspirate (72.2 ± 2.3 percent), and histologic analysis revealed high-density areas enriched with stromal cells (622.9 ± 145.6 percent) and extracellular matrix (871.2 ± 80.3 percent).
Stromal cell aggregates represent a biological agent that triplicates the cell density versus unprocessed lipoaspirate, low on oil and water fluids, and enriched extracellular matrix components.
再生细胞策略依赖于基质细胞植入物来获得可观察的临床效果。然而，确保治疗反应所需的有效细胞剂量尚不清楚。为了获得更高的细胞剂量，作者假设通过减少脂肪抽吸物中成熟脂肪细胞所占的体积，可以浓缩原始组织中存在的基质血管部分。
对人标准化脂肪抽吸物（n=6）进行离心（3 分钟，1200g），弃去水相。通过 2.4 和 1.2 毫米鲁尔到鲁尔转接器的剪切组织来实现机械分散。第二次离心（800g，10 分钟）后，将基质细胞聚集体与上清油相分离。通过其组成重量确定脂肪抽吸物的百分比组成。通过总 DNA 定量测量细胞含量，通过图像细胞计数测定部分细胞活力。通过苏木精和伊红以及 Masson 三色染色评估组织切片的组织学。
基质细胞聚集体将标准化脂肪抽吸物的质量减少到 28.6±4.2%。相应地，细胞密度增加了 222.6±63.3%（从 9.9±1.4×106 个细胞/g 增加到 31.3±6.6×106 个细胞/g；p<0.05）。与标准化脂肪抽吸物相比，基质细胞聚集体中的细胞活力无变化（71.3±2.5%），组织学分析显示富含基质细胞（622.9±145.6%）和细胞外基质（871.2±80.3%）的高密度区域。
基质细胞聚集体代表了一种生物制剂，与未处理的脂肪抽吸物相比，其细胞密度增加了三倍，油和水液体含量低，细胞外基质成分丰富。
