Institute of Pharmaceutical Science, King's College London, London, UK.
FASEB J. 2021 Jan;35(1):e21206. doi: 10.1096/fj.202001768RR.
Stem cell-derived exosomes are emerging as novel and clinically relevant cell-free therapeutics for regenerative therapy. This work focused on investigating the stimulation of fibroblasts by exosomes derived from umbilical cord-derived mesenchymal stem cells (ucMSC) in a defined serum-free three-dimensional (3D) culture. 3D culture of ucMSC was carried out in medium supplemented with KnockOut serum replacement (KO-medium) using the Aggrewell system. ucMSC in KO-medium formed spheroids with maintained size and integrity throughout culture. This enabled the isolation of vesicles from ucMSC spheroids in KO-medium with sizes that fall within the exosomal size range and were positive for the expression of canonical exosomal markers CD63, CD9, CD81, Alix, and TSG101. The ucMSC-derived exosomes (Exo ) were shown to significantly increase the migration and proliferation of murine fibroblasts in vitro. To conclude, 3D culture of ucMSC in defined serum-free KO-medium formed viable spheroids which enabled the isolation of Exo with the potential of accelerating wound healing.
干细胞衍生的外泌体作为新型的、具有临床相关性的细胞外治疗方法，正在成为再生治疗的研究热点。本研究专注于研究来自脐带间充质干细胞（ucMSC）的外泌体对成纤维细胞的刺激作用，该研究是在无血清的三维（3D）培养条件下进行的。ucMSC 在补充有 KnockOut 血清替代物（KO 培养基）的 Aggrewell 系统中进行 3D 培养。ucMSC 在 KO 培养基中形成的球体，在整个培养过程中保持大小和完整性不变。这使得可以从 KO 培养基中的 ucMSC 球体中分离出大小在细胞外体范围内的囊泡，并表达经典的细胞外体标志物 CD63、CD9、CD81、Alix 和 TSG101。ucMSC 衍生的外泌体（Exo）被证明可显著增加体外培养的小鼠成纤维细胞的迁移和增殖。综上所述，无血清的 KO 培养基中 3D 培养的 ucMSC 形成了有活力的球体，这使得能够分离具有加速伤口愈合潜力的 Exo。
