Kim Suna, Piao Jiyuan, Son Youngsook, Hong Hyun Sook
Department of Genetic Engineering, College of Life Science and Graduate School of Biotechnology, Kyung Hee University, Seochun-dong, Kiheung-ku, Yong In, 17104, South Korea.
Department of Genetic Engineering, College of Life Science and Graduate School of Biotechnology, Kyung Hee University, Seochun-dong, Kiheung-ku, Yong In, 17104, South Korea.
Biochem Biophys Res Commun. 2017 Mar 25;485(1):131-137. doi: 10.1016/j.bbrc.2017.02.036. Epub 2017 Feb 10.
Stem cells have tremendous promise to treat intractable diseases. Notably, adipose-derived stem cells (ADSCs) are actively being investigated because of ease of sampling and high repopulation capacity in vitro. ADSCs can exert a therapeutic effect through differentiation and paracrine potential, and these actions have been proven in many diseases, including cutaneous and inflammatory diseases. Transplantation of ADSCs necessitates therapeutic quantities and thus, long term ex vivo culture of ADSCs. However, this procedure can impair the activity of ADSCs and provoke cellular senescence, leading to low efficacy in vivo. Accordingly, strategies to restore cellular activity and inhibit senescence of stem cells during ex vivo culture are needed for stem cell-based therapies. This study evaluated a potential supplementary role of Substance P (SP) in ADSC ex vivo culture. After confirming that the ADSC cell cycle was damaged by passage 6 (p6), ADSCs at p6 were cultured with SP, and their proliferation rates, cumulative cell numbers, cytokine profiles, and impact on T/endothelial cells were assessed. Long-term culture weakened proliferation ability and secretion of the cytokines, transforming growth factor-beta 1 (TGF-beta1), vascular endothelial growth factor (VEGF), and stromal cell derived factor-1 alpha (SDF-1alpha) in ADSCs. However, SP treatment reduced the population doubling time (PDT), enabling gain of a sufficient number of ADSCs at early passages. In addition, SP restored cytokine secretion, enhancing the ADSC-mediated paracrine effect on T cell and human umbilical vein endothelial cells (HUVECs). Taken together, these results suggest that SP can retain the therapeutic effect of ADSCs by elevating their proliferative and paracrine potential in ex vivo culture.
干细胞在治疗难治性疾病方面具有巨大潜力。值得注意的是，脂肪来源干细胞（ADSCs）因其取样简便和体外高增殖能力而受到积极研究。ADSCs可通过分化和旁分泌潜能发挥治疗作用，并且这些作用已在包括皮肤和炎症性疾病在内的多种疾病中得到证实。ADSCs的移植需要治疗剂量，因此需要对ADSCs进行长期体外培养。然而，这一过程会损害ADSCs的活性并引发细胞衰老，导致体内疗效低下。因此，基于干细胞的治疗需要在体外培养过程中恢复细胞活性并抑制干细胞衰老的策略。本研究评估了P物质（SP）在ADSCs体外培养中的潜在补充作用。在确认第6代（p6）传代破坏了ADSC细胞周期后，将p6代的ADSCs与SP一起培养，并评估其增殖率、累积细胞数量、细胞因子谱以及对T/内皮细胞的影响。长期培养削弱了ADSCs的增殖能力以及细胞因子转化生长因子-β1（TGF-β1）、血管内皮生长因子（VEGF）和基质细胞衍生因子-1α（SDF-1α）的分泌。然而，SP处理缩短了群体倍增时间（PDT），使得在早期传代时能够获得足够数量的ADSCs。此外，SP恢复了细胞因子分泌，增强了ADSCs对T细胞和人脐静脉内皮细胞（HUVECs）的旁分泌作用。综上所述，这些结果表明SP可通过提高ADSCs在体外培养中的增殖和旁分泌潜能来保留其治疗效果。
