Department of Plastic, Reconstructive and Aesthetic Surgery, Geneva University Hospitals, Geneva, Switzerland.
Faculty of Medicine, Geneva University, Geneva, Switzerland.
Tissue Eng Part A. 2019 Nov;25(21-22):1550-1563. doi: 10.1089/ten.TEA.2018.0335. Epub 2019 May 21.
Nowadays autologous fibroblast application for skin repair presents an important clinical interest. In most cases skin cell culture is mandatory. However, cell expansion using xenogeneic or allogenic culture media presents some disadvantages, such as the risk of infection transmission or slow cell expansion. In this study, we investigated an autologous culture system to expand human skin fibroblast cells with the patient's own platelet-rich plasma (PRP). Human dermal fibroblasts were isolated from patients undergoing abdominoplasty, and blood was collected to prepare nonactivated PRP using the CuteCell™ PRP medical device. Cultures were followed up to 7 days using a medium supplemented with either fetal bovine serum (FBS) or PRP. Fibroblasts cultured in medium supplemented with PRP showed dose-dependently significantly higher proliferation rates (up to 7.7 times with 20% of PRP) and initiated a faster migration in the wound healing assay compared with FBS, while chromosomal stability was maintained. At high concentrations, PRP changed fibroblast morphology, inducing cytoskeleton rearrangement and an increase of alpha-smooth muscle actin and vimentin expression. Our findings show that autologous PRP is an efficient and cost-effective supplement for fibroblast culture, and should be considered as a safe alternative to xenogeneic/allogenic blood derivatives for cell expansion. Impact Statement Autologous dermal fibroblast graft is an important therapy in skin defect repair, but skin cell culture is mandatory in most cases. However, cell expansion using xenogeneic/allogenic culture media presents some disadvantages, such as the risk of infection transmission. We demonstrated that an autologous culture system with the patient's own platelet-rich plasma is an efficient, cost-effective, and safe supplement for fibroblast culture. As it respects the good manufacturing practices and regulatory agencies standards, it should be considered as a potent alternative and substitute to xenogeneic or allogenic blood derivatives for the validation of future clinical protocols using cell expansion.
如今,自体成纤维细胞在皮肤修复中的应用引起了重要的临床关注。在大多数情况下,皮肤细胞培养是必需的。然而,使用异种或同种培养介质进行细胞扩增存在一些缺点,例如感染传播的风险或细胞扩增缓慢。在这项研究中,我们研究了一种自体培养系统,该系统使用患者自身的富血小板血浆(PRP)来扩增人皮肤成纤维细胞。从接受腹部整形术的患者中分离出人真皮成纤维细胞,并采集血液使用 CuteCell™ PRP 医疗设备制备未激活的 PRP。使用补充有胎牛血清(FBS)或 PRP 的培养基对培养物进行 7 天的培养。与 FBS 相比,在 PRP 补充培养基中培养的成纤维细胞显示出剂量依赖性的显著更高的增殖率(高达 7.7 倍,PRP 为 20%),并且在伤口愈合测定中更快地迁移,同时保持染色体稳定性。在高浓度下,PRP 改变成纤维细胞形态,诱导细胞骨架重排,并增加α-平滑肌肌动蛋白和波形蛋白的表达。我们的研究结果表明,自体 PRP 是成纤维细胞培养的有效且经济有效的补充剂,并且应该被认为是异种/同种异体血液衍生物用于细胞扩增的安全替代品。
影响陈述
自体真皮成纤维细胞移植物是皮肤缺陷修复的重要疗法,但在大多数情况下,皮肤细胞培养是必需的。然而,使用异种/同种培养介质进行细胞扩增存在一些缺点,例如感染传播的风险。我们证明,使用患者自身的富血小板血浆的自体培养系统是成纤维细胞培养的有效、经济有效的且安全的补充剂。由于它符合良好生产规范和监管机构的标准,因此应被视为异种或同种异体血液衍生物的有效替代品和替代物,用于验证未来使用细胞扩增的临床方案。