Doctoral Programme in Biomedical Sciences, Faculty of Medicine, Brawijaya University, Malang, East Java, 65145, Indonesia.
Department of Cardiology and Vascular Medicine, Faculty of Medicine Universitas Airlangga, Dr. Soetomo Academic General Hospital, Surabaya, East Java, 60286, Indonesia.
F1000Res. 2024 Jun 11;11:851. doi: 10.12688/f1000research.122788.3. eCollection 2022.
There is an essential need for cardiomyocyte regeneration among patients with heart failure. Transplantation of dedifferentiated fat (DFAT) cells may lead to an improvement of cardiomyocyte regeneration among heart failure patients. We believe that DFAT cells are promising candidate cell sources for cardiac regeneration. However, the pathway underlying how DFAT cells of the adipose lineage differentiate into mature cardiomyocytes isn't fully understood.
We conducted an experimental laboratory study on isolated DFAT cells from adipose tissue of healthy adults. Then, we treated cells with different concentrations of reversine (10, 20 and 40 nM), and performed RNA extraction and cDNA synthesis. Next, we used a ceiling culture method based on the buoyancy properties of mature lipid-filled adipocytes. Stemness expression (Octamer-binding transcription factor 4 [Oct4], brachyury, Fetal liver kinase 1 [Flk-1]) was quantified by reverse transcription-quantitative (RT-q)PCR, while cardiomyocyte expression (Transcription factor GATA-4 [GATA4] and cardiac troponin T [cTnT]) was quantified by immunocytochemistry.
ANOVA with Tukey's post-hoc found that 10 nM reversine increased greater Flk-1 expression compared to the control group (MD: 5.037 + 0.998; < 0.001), but there were no significant changes among Oct4 (MD: 0.013 + 1.244; = 0.99) and brachyury expression (MD: 0.157 + 0.084; = 0.252). Kruskal-Wallis revealed that the expression of GATA4 (1.65 [0.41-1.98] to 0.015 [0.007-0.034]; 0.017) reduced significantly from day 7 until day 21 and cTnT (5.07 [6.62-8.91] to 8.22 [6.81-9.40]; = 0 increased significantly from day 7 until day 21.
Reversine could increase the expression of Flk-1, but it was unable to stimulate the expression of Oct4 and brachyury related to cell stemness. An optimal concentration of 10 nM reversine may have the greatest effect on enhancing the differentiation of DFAT cells into mature cardiomyocytes, as indicated by higher cTnT expression between cells.
心力衰竭患者需要心肌细胞再生。去分化脂肪（DFAT）细胞的移植可能会导致心力衰竭患者的心肌细胞再生得到改善。我们认为 DFAT 细胞是心脏再生有前途的候选细胞来源。然而，脂肪谱系的 DFAT 细胞如何分化为成熟心肌细胞的途径尚不完全清楚。
我们对来自健康成年人脂肪组织的分离的 DFAT 细胞进行了实验性实验室研究。然后，我们用不同浓度的雷维辛（10、20 和 40 nM）处理细胞，并进行 RNA 提取和 cDNA 合成。接下来，我们使用基于成熟含脂脂肪细胞浮力特性的天花板培养方法。通过逆转录定量（RT-q）PCR 定量干细胞表达（八聚体结合转录因子 4 [Oct4]、Brachyury、胎肝激酶 1 [Flk-1]），通过免疫细胞化学定量心肌细胞表达（转录因子 GATA-4 [GATA4]和肌钙蛋白 T [cTnT]）。
方差分析和 Tukey 事后检验发现，10 nM 雷维辛增加了 Flk-1 的表达，与对照组相比（MD：5.037 + 0.998；<0.001），但 Oct4 （MD：0.013 + 1.244；= 0.99）和 Brachyury 表达（MD：0.157 + 0.084；= 0.252）没有显著变化。Kruskal-Wallis 发现，GATA4 的表达（1.65 [0.41-1.98]至 0.015 [0.007-0.034]；0.017）从第 7 天到第 21 天显著降低，cTnT （5.07 [6.62-8.91]至 8.22 [6.81-9.40]；= 0）从第 7 天到第 21 天显著增加。
雷维辛可以增加 Flk-1 的表达，但不能刺激与细胞干性相关的 Oct4 和 Brachyury 的表达。10 nM 雷维辛的最佳浓度可能对增强 DFAT 细胞向成熟心肌细胞的分化有最大的影响，这表明细胞间的 cTnT 表达更高。
