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用标准化血小板裂解物补充培养的牙髓干细胞的高效修复行为

Highly Efficient Reparative Behaviour of Dental Pulp Stem Cells Cultured with Standardised Platelet Lysate Supplementation.

作者信息

Marrazzo Pasquale, Paduano Francesco, Palmieri Francesca, Marrelli Massimo, Tatullo Marco

机构信息

Tecnologica Research Institute, Biomedical Section, Crotone, Italy.

Tecnologica Research Institute, Biomedical Section, Crotone, Italy; Unit of Experimental Surgery, Calabrodental, Crotone, Italy.

出版信息

Stem Cells Int. 2016;2016:7230987. doi: 10.1155/2016/7230987. Epub 2016 Sep 28.

Abstract

Dental pulp is an accessible source of multipotent mesenchymal stromal cells (MSCs). The perspective role of dental pulp stem cells (DPSCs) in regenerative medicine demands an expansion and delivery which must deal with the safety issues about animal serum, usually required in cell culture practice. Human platelet lysate (PL) contains autologous growth factors and has been considered as valuable alternative to fetal bovine serum (FBS) in cell cultures. The optimum concentration to be added of such supplement is highly dependent on its preparation whose variability limits comparability of results. By experiments, we aimed to evaluate a standardised formulation of pooled PL. A low selected concentration of PL (1%) was able to support the growth and maintain the viability of the DPSCs. The use of PL in cell cultures did not impair cell surface signature typically expressed by MSCs and even upregulated the transcription of Sox2. Interestingly, DPSCs cultured in presence of PL exhibited a higher healing rate after injury and are less susceptible to toxicity mediated by exogenous HO than those cultured with FBS. Moreover, PL addition was shown as a suitable option for protocols promoting osteogenic and chondrogenic differentiation of DPSCs. Taken together, our results indicated that PL is a valid substitute of FBS to culture and differentiate DPSCs for clinical-grade use.

摘要

牙髓是多能间充质基质细胞(MSCs)的一个可获取来源。牙髓干细胞(DPSCs)在再生医学中的潜在作用要求进行细胞扩增和递送,而这必须解决细胞培养实践中通常所需的动物血清的安全性问题。人血小板裂解物(PL)含有自体生长因子,在细胞培养中被认为是胎牛血清(FBS)的有价值替代品。此类补充剂的最佳添加浓度高度依赖于其制备方法,其变异性限制了结果的可比性。通过实验,我们旨在评估一种标准化的混合PL配方。低浓度(1%)的PL能够支持DPSCs的生长并维持其活力。在细胞培养中使用PL不会损害MSCs通常表达的细胞表面特征,甚至上调了Sox2的转录。有趣的是,与用FBS培养的DPSCs相比,在PL存在下培养的DPSCs在损伤后表现出更高的愈合率,并且对外源性HO介导的毒性更不敏感。此外,添加PL被证明是促进DPSCs成骨和成软骨分化方案的合适选择。综上所述,我们的结果表明,PL是用于临床级培养和分化DPSCs的FBS的有效替代品。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b837/5059612/000bbc5197e6/SCI2016-7230987.001.jpg

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