Impact of cell culture parameters on production and vascularization bioactivity of mesenchymal stem cell-derived extracellular vesicles.
Impact of cell culture parameters on production and vascularization bioactivity of mesenchymal stem cell-derived extracellular vesicles.
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DOI：
10.1002/btm2.10065
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发表时间：
2017-06
影响因子：
7.4
通讯作者：
Jay SM
中科院分区：
文献类型：
--
作者：
Patel DB;Gray KM;Santharam Y;Lamichhane TN;Stroka KM;Jay SM
间充质干细胞（MSC）衍生的细胞外囊泡（EV）已成为许多应用的潜在治疗剂。EV在安全性、稳定性和清除率方面比基于细胞的疗法具有潜在优势，但必须解决生产和效力限制，以实现基于EV的方法的最终转化。因此，我们试图研究特定细胞培养参数对MSC EV生产和生物活性的作用，以告知可扩展EV生物制造的合理设计参数。我们报告了显著降低的MSC EV血管化生物活性，如通过内皮细胞间隙闭合测定所测量的，随着通过胰蛋白酶消化培养的传代次数的增加，特别是超过第4代。我们进一步表明，增加EV收集频率在24小时内从相同的初始数量的MSC产生更高数量的EV。最后，我们证明了培养瓶中细胞接种密度的降低导致MSC和其他细胞类型中每个细胞的EV产量增加。总体而言，这些研究强调了在实验室规模生产治疗性EV时仔细考虑细胞传代次数和细胞接种密度参数以及合理设计大规模EV生物制造方案的必要性。
Mesenchymal stem cell (MSC)‐derived extracellular vesicles (EVs) have emerged as potential therapeutic agents for numerous applications. EVs offer potential advantages over cell‐based therapies with regard to safety, stability and clearance profiles, however production and potency limitations must be addressed to enable eventual translation of EV‐based approaches. Thus, we sought to examine the role of specific cell culture parameters on MSC EV production and bioactivity toward informing rational design parameters for scalable EV biomanufacturing. We report significantly reduced MSC EV vascularization bioactivity, as measured by an endothelial cell gap closure assay, with increasing passage in culture by trypsinization, especially beyond passage 4. We further show that increased frequency of EV collection yielded higher numbers of EVs from the same initial number of MSCs over a 24 hr period. Finally, we demonstrate that decreased cell seeding density in culture flasks resulted in increased production of EVs per cell in MSCs and other cell types. Overall, these studies highlight the need for careful consideration of the parameters of cell passage number and cell seeding density in the production of therapeutic EVs at laboratory scale and for rational design of large‐scale EV biomanufacturing schemes.
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影响因子：
3.7
作者：
Li J;Chen X;Yi J;Liu Y;Li D;Wang J;Hou D;Jiang X;Zhang J;Wang J;Zen K;Yang F;Zhang CY;Zhang Y
通讯作者：
Zhang Y
影响因子：
2.8
作者：
Lamichhane, Tek N.;Jeyaram, Anjana;Patel, Divya B.;Parajuli, Babita;Livingston, Natalie K.;Arumugasaamy, Navein;Schardt, John S.;Jay, Steven M.
通讯作者：
Jay, Steven M.
影响因子：
14
作者：
Chung, Cindy;Beecham, Michael;Mauck, Robert L.;Burdick, Jason A.
通讯作者：
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影响因子：
4.1
作者：
Abdeen, Amr A.;Weiss, Jared B.;Kilian, Kristopher A.
通讯作者：
Kilian, Kristopher A.
影响因子：
7.4
作者：
Chen TS;Arslan F;Yin Y;Tan SS;Lai RC;Choo AB;Padmanabhan J;Lee CN;de Kleijn DP;Lim SK
通讯作者：
Lim SK