富血小板纤维蛋白基质(PRFM)中间充质干细胞组织再生的相容性分析

G. Pratama, B. Wiweko, N. Sandora, Evanti Kusumawardhani, D. Rahayu, Kamila Puspita, M. Reksodiputro
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引用次数: 2

摘要

间充质干细胞(MSCs)是一种能够分化为各种间充质或非间充质细胞系的祖细胞。在大多数组织的愈合和再生过程中,间充质干细胞也对损伤起作用。富血小板血浆(PRP)已被报道可诱导子宫内膜再生,因此研究了一种致密形式的PRP,即富血小板纤维蛋白基质(PRFM),其承载MSC的能力以及MSC在基质中的行为。加入25mm的CaCl2,形成直径为5cm的硬币形状,形成PRP凝胶。将每个基质切成两半,直接播种骨髓(hBM-MSC)和脐带MSC (hUC-MSC),播种密度为2000细胞。cm−2。H&E染色分析初始细胞附着,MTT法定量评估细胞活力,Live/Dead染色定性评估细胞活力。与hBM-MSC相比,hUC-MSC在PRFM中表现出最高的活力,特别是在第1天和第5天的孵育期间。孵育10 ~ 15天后,存活率下降。活/死染色表明细胞群过于拥挤,大部分细胞死亡。这可能是由于窒息。随着时间的推移,hUC-MSC和hBM-MSC可以增殖并浸润PRFM,尽管在孵育后期死亡细胞的比例更占优势。PRFM可用于将MSC输送到损伤部位。然而,直接播种法在前24小时的孵育中并不是最理想的方法,因为它们很容易分离。
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Cito-compability analysis of mesenchymal stem cells in platelet rich fibrin matrix (PRFM) for tissue regeneration
Mesenchymal stem cells (MSCs) is progenitor cell that capable to differentiate to various mesenchymal or non-mesenchymal lineages. MSC also play a role towards injury during healing and regeneration processes in most tissue. Platelet rich plasma (PRP) had been reported to induce endometrium regeneration, therefore a compacted form of PRP, known as platelet rich fibrin matrix (PRFM), was studied for its capability to host MSC and the behavior of MSC in the matrix. PRP gelation was generated by adding 25 mM of CaCl2 to form a coin shape with diameter of 5 cm. Each matrix was cut into half, seeded directly with bone marrow (hBM-MSC) and umbilical cord MSC (hUC-MSC) with seeding density of 2,000 cell.cm−2. Initial cellular attachment was analyzed using H&E staining, while the cellular viability was assessed quantitatively using MTT assay and qualitatively using Live/Dead staining. hUC-MSC showed the highest viability compared to hBM-MSC when delivered in PRFM, especially during the first and five days incubation. After 10 dan 15 days incubation, the viability had dropped. The Live/Dead staining indicated too crowded population and most cells were dead. This might be due to asphyxia. hUC-MSC and hBM-MSC were seen to proliferated and infiltrated the PRFM over the time, although the ratio of dead cells was more dominant in the later day of incubation. PRFM can be used to deliver MSC to the site injury. However the direct seeding method was not optimal to hold the cells on place during the first 24 h incubation because they were easily detached.
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