Echeverry Santiago, Grismaldo Adriana, Sánchez Charles, Sierra Cristian, Henao Juan C, Granados Sara T, Sutachán Jhon-Jairo, Torres Yolima P
Departamento de Nutrición y Bioquímica, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, Colombia.
Front Physiol. 2020 Mar 24;11:210. doi: 10.3389/fphys.2020.00210. eCollection 2020.
Due to their capacity to proliferate, migrate, and differentiate, mesenchymal stem cells (MSCs) are considered to be good candidates for regenerative medicine applications. The mechanisms underlying proliferation and differentiation of MSCs have been studied. However, much less is known about the mechanisms regulating the migration of MSCs. Platelet lysate (PL), a supplement used to promote cell expansion, has been shown to promote MSCs migration; however, the underlying mechanism are unknown. Here, by using adipose-derived rat MSCs (rMSCs) and the scratch assay in the absence and presence of various BK channels modulators, we evaluated the role of BK channels in mediating the PL-stimulated migration of rMSCs. We found that 5% PL increased rMSCs migration, and this effect was blocked by the addition of the BK channel selective antagonist Iberiotoxin (IBTX). In the absence of PL, the BK channel agonist NS1619, stimulated rMSCs migration to similar level as 5% PL. Addition of both NS1619 and 5% PL resulted in an increase in rMSCs migration, that was higher than when either one was added individually. From whole-cell recordings, it was found that the addition of 5% PL increased the magnitude of BK current density. By using Western blot and flow cytometry, it was found that PL did not affect the expression of BK channels. Together, our results indicate that as shown in other cell types, activation of BK channels by themselves also promote rMSC migration, and show that activation of BK channels contribute to the observed PL-induced increase in migration of rMSC.
由于间充质干细胞（MSCs）具有增殖、迁移和分化的能力，它们被认为是再生医学应用的良好候选者。MSCs增殖和分化的潜在机制已得到研究。然而，关于调节MSCs迁移的机制却知之甚少。血小板裂解物（PL）是一种用于促进细胞扩增的补充剂，已被证明可促进MSCs迁移；然而，其潜在机制尚不清楚。在这里，我们使用脂肪来源的大鼠间充质干细胞（rMSCs），并在存在和不存在各种BK通道调节剂的情况下进行划痕试验，评估BK通道在介导PL刺激的rMSCs迁移中的作用。我们发现5%的PL增加了rMSCs的迁移，并且这种作用被BK通道选择性拮抗剂iberiotoxin（IBTX）的添加所阻断。在没有PL的情况下，BK通道激动剂NS1619刺激rMSCs迁移到与5%PL相似的水平。同时添加NS1619和5%PL导致rMSCs迁移增加，这比单独添加任何一种时都要高。通过全细胞记录发现，添加5%PL增加了BK电流密度的幅度。通过蛋白质印迹和流式细胞术发现，PL不影响BK通道的表达。总之，我们的结果表明，与其他细胞类型一样，BK通道自身的激活也促进rMSC迁移，并表明BK通道的激活有助于观察到的PL诱导的rMSC迁移增加。
