Terumo BCT®, Inc., Lakewood, CO, USA.
Terumo BCT®, Inc., Lakewood, CO, USA.
Mater Sci Eng C Mater Biol Appl. 2019 Mar;96:77-85. doi: 10.1016/j.msec.2018.10.081. Epub 2018 Oct 26.
The addition of a coating reagent to promote cell adherence is necessary to prepare the membrane surface of the Quantum® Cell Expansion System hollow-fiber bioreactor for the culture of mesenchymal stem cells. In this study, the efficacy of 8 potential coating reagents has been compared in terms of the doubling times of their cell populations, cell morphology, characterization via flow cytometry, and capacity for trilineage differentiation. Human fibronectin (FN), pooled human cryoprecipitate (CPPT), and recombinant human vitronectin (VN) were successful as coating reagents, and each product has advantages in different cell culture contexts. Mesenchymal stem cells harvested from Quantum cultured with each of these 3 compounds as coating reagents all met International Society for Cellular Therapy standards for plastic adherence, surface marker expression, and successful trilineage differentiation. No significant differences were observed among the doubling times from Quantum harvests using FN, CPPT, or VN as coating reagents (P = 0.31). Coating with gelatin, human serum albumin, collagen I, poly‑l‑lysine, and poly‑d‑lysine resulted in significantly lower harvest yield; these agents are not recommended for use as coating reagents in the Quantum system.
为了准备 Quantum®细胞扩增系统中空纤维生物反应器的膜表面用于间充质干细胞的培养,需要添加涂层试剂以促进细胞黏附。在这项研究中,比较了 8 种潜在的涂层试剂在细胞群体倍增时间、细胞形态、流式细胞术特征以及三系分化能力方面的效果。人纤维连接蛋白(FN)、混合人冷沉淀(CPPT)和重组人 vitronectin(VN)作为涂层试剂是成功的,每种产品在不同的细胞培养环境中都有优势。用这 3 种化合物作为涂层试剂培养的 Quantum 收获的间充质干细胞均符合国际细胞治疗学会对塑料黏附、表面标志物表达和成功三系分化的标准。使用 FN、CPPT 或 VN 作为涂层试剂时,从 Quantum 收获的倍增时间没有观察到显著差异(P = 0.31)。用明胶、人血清白蛋白、胶原 I、聚赖氨酸和聚天冬氨酸作为涂层试剂,收获产量明显较低;不建议将这些试剂用于 Quantum 系统中的涂层试剂。