Department of Plastic and Hand Surgery, Laboratory for Tissue Engineering and Regenerative Medicine, University Hospital of Erlangen, Friedrich-Alexander University of Erlangen-Nürnberg (FAU), 91054, Erlangen, Germany.
Department of Plastic Surgery, Hand Surgery - Burn Center, University Hospital RWTH Aachen, 52074, Aachen, Germany.
BMC Mol Cell Biol. 2023 May 15;24(1):18. doi: 10.1186/s12860-023-00478-1.
For the purpose of skeletal muscle engineering, primary myoblasts (Mb) and adipogenic mesenchymal stem cells (ADSC) can be co-cultured and myogenically differentiated. Electrospun composite nanofiber scaffolds represent suitable matrices for tissue engineering of skeletal muscle, combining both biocompatibility and stability Although growth differentiation factor 11 (GDF11) has been proposed as a rejuvenating circulating factor, restoring skeletal muscle function in aging mice, some studies have also described a harming effect of GDF11. Therefore, the aim of the study was to analyze the effect of GDF11 on co-cultures of Mb and ADSC on poly-ε-caprolactone (PCL)-collagen I-polyethylene oxide (PEO)-nanofibers.
Human Mb were co-cultured with ADSC two-dimensionally (2D) as monolayers or three-dimensionally (3D) on aligned PCL-collagen I-PEO-nanofibers. Differentiation media were either serum-free with or without GDF11, or serum containing as in a conventional differentiation medium. Cell viability was higher after conventional myogenic differentiation compared to serum-free and serum-free + GDF11 differentiation as was creatine kinase activity. Immunofluorescence staining showed myosine heavy chain expression in all groups after 28 days of differentiation without any clear evidence of more or less pronounced expression in either group. Gene expression of myosine heavy chain (MYH2) increased after serum-free + GDF11 stimulation compared to serum-free stimulation alone.
This is the first study analyzing the effect of GDF11 on myogenic differentiation of Mb and ADSC co-cultures under serum-free conditions. The results of this study show that PCL-collagen I-PEO-nanofibers represent a suitable matrix for 3D myogenic differentiation of Mb and ADSC. In this context, GDF11 seems to promote myogenic differentiation of Mb and ADSC co-cultures compared to serum-free differentiation without any evidence of a harming effect.
为了进行骨骼肌工程，原代成肌细胞（Mb）和脂肪间充质干细胞（ADSC）可以共培养并进行肌源性分化。静电纺丝复合纳米纤维支架是一种适合骨骼肌组织工程的基质，具有生物相容性和稳定性。虽然生长分化因子 11（GDF11）已被提出作为一种恢复衰老小鼠骨骼肌功能的再生循环因子，但一些研究也描述了 GDF11 的有害作用。因此，本研究旨在分析 GDF11 对 Mb 和 ADSC 共培养物在聚己内酯（PCL）-胶原蛋白 I-聚氧化乙烯（PEO）-纳米纤维上的影响。
将人 Mb 与 ADSC 二维（2D）共培养为单层或三维（3D）共培养于排列的 PCL-胶原蛋白 I-PEO-纳米纤维上。分化培养基为无血清或含血清，无血清或无血清+GDF11，或含血清，如常规分化培养基。与无血清和无血清+GDF11 分化相比，常规肌源性分化后的细胞活力更高，肌酸激酶活性也更高。免疫荧光染色显示，所有组在分化 28 天后均有肌球蛋白重链表达，而在任何一组中均无更明显或更不明显的表达。与无血清刺激相比，无血清+GDF11 刺激后肌球蛋白重链（MYH2）的基因表达增加。
这是第一项分析 GDF11 在无血清条件下对 Mb 和 ADSC 共培养物肌源性分化影响的研究。本研究结果表明，PCL-胶原蛋白 I-PEO-纳米纤维是 Mb 和 ADSC 三维肌源性分化的合适基质。在这种情况下，与无血清分化相比，GDF11 似乎促进了 Mb 和 ADSC 共培养物的肌源性分化，而没有任何有害作用的证据。
