Endothelial Cells Increase Mesenchymal Stem Cell Differentiation in Scaffold-Free 3D Vascular Tissue.

IF 3.5 3区 医学 Q3 CELL & TISSUE ENGINEERING Tissue Engineering Part A Pub Date : 2024-09-12 DOI:10.1089/ten.TEA.2024.0122
William G DeMaria, Andre E Figueroa-Milla, Abigail Kaija, Anne E Harrington, Benjamin Tero, Larisa Ryzhova, Lucy Liaw, Marsha W Rolle
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Abstract

In this study, we present a versatile, scaffold-free approach to create ring-shaped engineered vascular tissue segments using human mesenchymal stem cell-derived smooth muscle cells (hMSC-SMCs) and endothelial cells (ECs). We hypothesized that incorporation of ECs would increase hMSC-SMC differentiation without compromising tissue ring strength or fusion to form tissue tubes. Undifferentiated hMSCs and ECs were co-seeded into custom ring-shaped agarose wells using four different concentrations of ECs: 0%, 10%, 20%, and 30%. Co-seeded EC and hMSC rings were cultured in SMC differentiation medium for a total of 22 days. Tissue rings were then harvested for histology, Western blotting, wire myography, and uniaxial tensile testing to examine their structural and functional properties. Differentiated hMSC tissue rings comprising 20% and 30% ECs exhibited significantly greater SMC contractile protein expression, endothelin-1 (ET-1)-meditated contraction, and force at failure compared with the 0% EC rings. On average, the 0%, 10%, 20%, and 30% EC rings exhibited a contractile force of 0.745 ± 0.117, 0.830 ± 0.358, 1.31 ± 0.353, and 1.67 ± 0.351 mN (mean ± standard deviation [SD]) in response to ET-1, respectively. Additionally, the mean maximum force at failure for the 0%, 10%, 20%, and 30% EC rings was 88.5 ± 36. , 121 ± 59.1, 147 ± 43.1, and 206 ±  0.8 mN (mean ± SD), respectively. Based on these results, 30% EC rings were fused together to form tissue-engineered blood vessels (TEBVs) and compared with 0% EC TEBV controls. The addition of 30% ECs in TEBVs did not affect ring fusion but did result in significantly greater SMC protein expression (calponin and smoothelin). In summary, co-seeding hMSCs with ECs to form tissue rings resulted in greater contraction, strength, and hMSC-SMC differentiation compared with hMSCs alone and indicates a method to create a functional 3D human vascular cell coculture model.

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内皮细胞可促进无支架三维血管组织中间充质干细胞的分化。
在这项研究中,我们提出了一种多功能、无支架的方法,利用人间充质干细胞衍生的平滑肌细胞(hMSC-SMC)和内皮细胞(EC)创建环形工程血管组织片段。我们假设,EC的加入将增加hMSC-SMC的分化,而不会影响组织环的强度或融合形成组织管。将未分化的 hMSCs 和 ECs 共同播种到定制的环形琼脂糖孔中,使用四种不同浓度的 ECs:0、10、20 和 30%。共种的 EC 和 hMSC 环在 SMC 分化培养基中总共培养了 22 天。然后取组织环进行组织学、Western 印迹、金属丝肌电图和单轴拉伸测试,以检查其结构和功能特性。与含 20% 和 30% EC 的组织环相比,含 20% 和 30% EC 的分化 hMSC 组织环在 SMC 收缩蛋白表达、内皮素-1(ET-1)诱导的收缩和失效时的力量方面均明显高于含 0% EC 的组织环。平均而言,0、10、20 和 30% EC 环对 ET-1 的反应收缩力分别为 0.745 ± 0.117、0.830 ± 0.358、1.31 ± 0.353 和 1.67 ± 0.351 mN(平均值 ± SD)。此外,0、10、20 和 30% EC 环的平均最大破坏力分别为 88.5 ± 36.2、121 ± 59.1、147 ± 43.1 和 206 ± 20.8 mN(平均值 ± SD)。基于这些结果,30% EC 环被融合在一起形成组织工程血管(TEBV),并与 0% EC TEBV 对照组进行比较。在 TEBV 中添加 30% 的 EC 不会影响环的融合,但会导致 SMC 蛋白表达(钙蛋白和平滑肌蛋白)显著增加。总之,与单独使用 hMSCs 相比,将 hMSCs 与 ECs 共同接种形成组织环会产生更大的收缩力、强度和 hMSC-SMC 分化,这表明有一种方法可以创建功能性三维人类血管细胞共培养模型。
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来源期刊
Tissue Engineering Part A
Tissue Engineering Part A Chemical Engineering-Bioengineering
CiteScore
9.20
自引率
2.40%
发文量
163
审稿时长
3 months
期刊介绍: Tissue Engineering is the preeminent, biomedical journal advancing the field with cutting-edge research and applications that repair or regenerate portions or whole tissues. This multidisciplinary journal brings together the principles of engineering and life sciences in the creation of artificial tissues and regenerative medicine. Tissue Engineering is divided into three parts, providing a central forum for groundbreaking scientific research and developments of clinical applications from leading experts in the field that will enable the functional replacement of tissues.
期刊最新文献
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