Department of Obstetrics and Gynecology, University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany.
BiomaTiCS-Biomaterials, Tissues, and Cells in Science, University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany.
Biomed Res Int. 2022 Mar 3;2022:3807314. doi: 10.1155/2022/3807314. eCollection 2022.
Fetal calf serum (FCS) is used for cell culture, as it provides the cells with various growth-promoting compounds. For applications in humans, FCS does not meet the required safety standards and should be replaced by an appropriate substitute. This study analyzed the suitability of using human platelet lysate (hPL) as a substitute for FCS in endothelial cell cultures for and tissue engineering applications. The focus was placed on standardized, commercially available hPLs (MultiPL'30, MultiPL'100), which are approved for applications in humans, and compared to laboratory-prepared hPLs (lp-hLP). Human umbilical vein endothelial cells (HUVEC) were cultured with FCS or with different hPLs. Cell morphology, proliferation, viability, apoptosis, and necrosis, as well as the organization of vascular structures, were assessed. No morphological changes were noticed when FCS was replaced by standardized hPLs in concentrations of 1-10%. In contrast, the use of lp-hLPs led to irregular cell shape and increased vacuolization of the cytoplasm. HUVEC proliferation and viability were not compromised by using media supplemented with standardized hPLs or pl-hPLs in concentrations of 1-10%, compared to cells grown in media supplemented with 20% FCS. The apoptosis rate using lp-hPLs was higher compared to the use of standardized hPLs. The necrosis rate tended to be lower when FCS was replaced by hPLs. HUVEC formed more pronounced capillary-like structures when the media were supplemented with hPLs instead of supplementation with FCS. Thus, compared to the use of FCS, the use of hPLs was beneficial for the growth and optimal expression of functional endothelial cell characteristics during experiments. Commercially available hPLs proved to be particularly suitable, as they led to reproducible results during experiments, while meeting the safety requirements for in vivo use.
胎牛血清(FCS)用于细胞培养,因为它为细胞提供了各种促进生长的化合物。对于人类应用,FCS 不符合所需的安全标准,应被适当的替代品取代。本研究分析了用人血小板裂解物(hPL)替代 FCS 在用于和组织工程应用的内皮细胞培养中的适用性。重点放在了标准化的、商业上可用的 hPL(MultiPL'30、MultiPL'100)上,这些 hPL 已被批准用于人类应用,并与实验室制备的 hPL(lp-hLP)进行了比较。用人脐静脉内皮细胞(HUVEC)在 FCS 或不同 hPL 中培养。评估了细胞形态、增殖、活力、凋亡和坏死以及血管结构的组织。当 FCS 被标准化的 hPL 以 1-10%的浓度替代时,没有观察到形态变化。相比之下,使用 lp-hLPs 导致细胞质不规则形状和空泡化增加。与在含有 20%FCS 的培养基中生长的细胞相比,使用浓度为 1-10%的标准化 hPL 或 lp-hPL 补充的培养基不会损害 HUVEC 的增殖和活力。与使用标准化 hPL 相比,使用 lp-hPLs 的细胞凋亡率更高。当 FCS 被 hPL 替代时,细胞坏死率有降低的趋势。与补充 FCS 相比,当培养基中补充 hPL 时,HUVEC 形成更明显的毛细血管样结构。因此,与使用 FCS 相比,在实验中使用 hPL 有利于生长和最佳表达功能性内皮细胞特征。商业上可用的 hPL 被证明是特别合适的,因为它们在实验中产生了可重复的结果,同时满足了体内应用的安全要求。