Chouw Angliana, Sartika Cynthia Retna, Milanda Tiana, Faried Ahmad
Doctoral Program, Faculty of Pharmacy Universitas Padjadjaran, Jatinangor, West Java, Indonesia.
Research and Development, Prodia StemCell Indonesia, Jakarta, DKI, Jakarta, Indonesia.
Stem Cells Cloning. 2022 Apr 14;15:1-9. doi: 10.2147/SCCAA.S356763. eCollection 2022.
Umbilical cord mesenchymal stem cells (UC-MSCs)-derived secretome is currently used in regenerative therapy. MSCs are believed to secrete a wide spectrum of bioactive molecules which give paracrine effects in immunomodulation and regenerative capacities. One group that was found in secretome is interleukins (ILs), a cytokine that plays an essential role in the process of proliferation, differentiation, maturation, migration, and adhesion of immune cells. However, as there are many types of ILs, the profile of ILs in the UC-MSCs-derived secretome has been limitedly reported. Therefore, in this study, we would like to profile and detect the interleukin concentration secreted by UC-MSCs.
UC-MSCs-derived secretome was collected from UC-MSCs passage 5 after 24- and 48-hour incubation (n=9). Secretome was filtered using 0.2 µm and stored at -80°C for further detection. All samples were normalized before the interleukin (IL-2, IL-4, IL-6, IL-9, IL-10, IL-12, IL-17A) detection using a MACSPlex Cytokine Kit.
The IL-6 has the highest concentration among other interleukins in both groups and increases significantly (<0.003) after incubation for 48 hours. The pro-inflammatory factors are decreasing while anti-inflammatory factors are increasing after 48-hour incubation.
Our studies show that the UC-MSCs secrete pro- and anti-inflammatory interleukins. The concentration of anti-inflammatory interleukins shows to be increasing, while the pro-inflammatory interleukins are decreasing within the longer incubation time, but this not be applicable for IL-10 and IL-6. IL-6 has the highest concentration among other ILs. These results may provide important clues regarding when is the right time for secretome to be used in therapy patients, because all the molecules in the secretome can lead to many clinical manifestations.
脐带间充质干细胞（UC-MSCs）分泌组目前用于再生治疗。人们认为间充质干细胞分泌多种生物活性分子，这些分子在免疫调节和再生能力方面具有旁分泌作用。在分泌组中发现的一类物质是白细胞介素（ILs），这是一种在免疫细胞增殖、分化、成熟、迁移和黏附过程中起重要作用的细胞因子。然而，由于白细胞介素种类繁多，关于UC-MSCs分泌组中白细胞介素的谱型报道有限。因此，在本研究中，我们希望分析并检测UC-MSCs分泌的白细胞介素浓度。
在培养24小时和48小时后，从第5代UC-MSCs收集UC-MSCs分泌组（n = 9）。分泌组用0.2 µm滤器过滤，并储存在-80°C以备进一步检测。在使用MACSPlex细胞因子试剂盒检测白细胞介素（IL-2、IL-4、IL-6、IL-9、IL-10、IL-12、IL-17A）之前，对所有样品进行标准化处理。
在两组中，IL-6在其他白细胞介素中浓度最高，培养48小时后显著增加（<0.003）。培养48小时后，促炎因子减少，而抗炎因子增加。
我们的研究表明，UC-MSCs分泌促炎和抗炎白细胞介素。在较长的培养时间内，抗炎白细胞介素的浓度呈上升趋势，而促炎白细胞介素的浓度呈下降趋势，但IL-10和IL-6不适用。IL-6在其他白细胞介素中浓度最高。这些结果可能为分泌组用于治疗患者的最佳时机提供重要线索，因为分泌组中的所有分子都可能导致许多临床表现。
