Introduction

Cardiovascular disease is the second killer across the globe, while coronary disease is the major cause. Cell therapy is one alternative to regenerate the infarcted heart wall.

Materials and Methods

In this study, the cardiomyogenesis capacity of human adipose stem cells (hAdSC) and human cardiomyocytes (hCardio) cultured in a 3-D biological scaffold (decellularised amnion bilayer) for nine days in a static condition was investigated. The cardiomyogenesis capacity of hAdSC were identified using immunohistochemistry and RT-PCR. The population of the cells isolated from the heart tissue expressed cTnT-1 (13.38 ± 11.38%), cKit (7.85 ± 4.2%), ICAM (85.53 ± 8.69%), PECAM (61.63 ± 7.18%) and VCAM (35.9 ± 9.11%), while from the fat tissue expressed the mesenchymal phenotypes (CD73, CD90, CD105, but not CD45, CD34, CD11b, CD19 and HLA-DR). Two age groups of hAdSC donors were compared, the youngsters (30-40yo) and the elderly (60–70 yo).

Results

The co-culture showed that after 5-day incubation, the seeded graft in the hAdSC-30 group had a tube-like appearance while the hAdSC-60 group demonstrated a disorganised pattern, despite of the MSC expressions of the hAdSC-60 were significantly higher. Initial co-culture showed no difference of ATP counts among all groups, however the hAdSC-30 group had the highest ATP count after 9 days culture (p = 0.004). After normalising to the normal myocardium, only the hAdSC-60 group expressed cTnT and MHC, very low, seen during the initial cultivation, but then disappeared. Meanwhile, the hAdSC-30 group expressed α-actinin, MHC and cTnT in the Day-5. The PPAR also was higher in the Day-5 compared to the Day-9 (p < 0.005).

Conclusion

Cardiomyogenesis capacity of hAdSC co-cultured with hCardio in a 3-D scaffold taken from the 30–40yo donor showed better morphology and viability than the 60-70yo group, but maintained less than 5 days in this system.

中文翻译：

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细胞种子心脏贴片的体外制备；羊膜双层贴片中脂肪来源的间充质干细胞和心肌细胞的共培养

介绍

心血管疾病是全球第二大杀手，而冠状动脉疾病是主要原因。细胞疗法是再生梗塞心壁的一种替代方法。

材料和方法

在这项研究中，研究了在静态条件下在 3-D 生物支架（脱细胞羊膜双层）中培养 9 天的人脂肪干细胞 (hAdSC) 和人心肌细胞 (hCardio) 的心肌生成能力。使用免疫组织化学和 RT-PCR 鉴定了 hAdSC 的心肌生成能力。从心脏组织中分离出的细胞群表达 cTnT-1 (13.38 ± 11.38%)、cKit (7.85 ± 4.2%)、ICAM (85.53 ± 8.69%)、PECAM (61.63 ± 7.18%) 和 VCAM (35.9 ± 9.11) %)，而从脂肪组织表达间充质表型（CD73、CD90、CD105，但不包括 CD45、CD34、CD11b、CD19 和 HLA-DR）。比较了两个年龄组的 hAdSC 供体，年轻人（30-40 岁）和老年人（60-70 岁）。

结果

共培养结果显示，hAdSC-30 组接种后的移植物在孵育 5 天后呈管状外观，而 hAdSC-60 组则表现出杂乱无章的模式，尽管 hAdSC-60 的 MSC 表达显着。更高。最初的共培养显示所有组之间的 ATP 计数没有差异，但在培养 9 天后，hAdSC-30 组的 ATP 计数最高（p  = 0.004）。归一化至正常心肌后，只有hAdSC-60组表达cTnT和MHC，非常低，在初始培养时可见，但随后消失。同时，hAdSC-30 组在第 5 天表达 α-actinin、MHC 和 cTnT。与第 9 天相比，第 5 天的 PPAR 也更高（p < 0.005）。

结论

在取自 30-40yo 供体的 3-D 支架中，hAdSC 与 hCardio 共培养的心肌生成能力显示出比 60-70yo 组更好的形态和活力，但在该系统中维持不到 5 天。