Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons.
Engineering skeletal muscle tissues with advanced maturity improves synapse formation with human induced pluripotent stem cell-derived motor neurons.
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DOI：
10.1063/5.0054984
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发表时间：
2021-09
影响因子：
6
通讯作者：
McCain ML
中科院分区：
文献类型：
--
作者：
Santoso JW;Li X;Gupta D;Suh GC;Hendricks E;Lin S;Perry S;Ichida JK;Dickman D;McCain ML
为了开发神经肌肉疾病的有效治疗方法，迫切需要与人类相关的神经肌肉组织体外模型，以在细胞和分子水平上探索疾病机制。然而，以前的尝试共培养运动神经元和骨骼肌导致相对不成熟的神经肌肉接头（NMJ）。在这项研究中，通过优化共培养肌肉组织的成熟度来改善由人诱导多能干细胞（hiPSC）衍生的运动神经元形成的NMJ。首先，比较了在微模塑明胶水凝胶上从C2C12小鼠成肌细胞系、冷冻保存的原代人成肌细胞和新鲜分离的原代鸡成肌细胞工程化的肌肉组织。三周后，只有鸡肌肉组织保持稳定地粘附到水凝胶上，并表现出生肌指数和应力产生的逐渐增加，接近天然肌肉组织产生的值。在与hiPSC衍生的运动神经元共培养三周后，工程化的鸡肌肉组织形成了NMJ，突触前和突触后标记物的共定位增加，以及突触活动的频率和幅度增加，超过了先前体外模型的结构和功能成熟度。工程鸡肌肉组织也表现出随着时间的推移与肌节成熟和神经支配相关的基因表达增加，揭示了可能有助于增强NMJ形成的分子途径的新见解。这些用于工程化具有相对成熟的NMJ的高级神经肌肉组织并询问其结构和功能的方法在神经肌肉疾病建模和药物开发中具有许多应用。
To develop effective cures for neuromuscular diseases, human-relevant in vitro models of neuromuscular tissues are critically needed to probe disease mechanisms on a cellular and molecular level. However, previous attempts to co-culture motor neurons and skeletal muscle have resulted in relatively immature neuromuscular junctions (NMJs). In this study, NMJs formed by human induced pluripotent stem cell (hiPSC)-derived motor neurons were improved by optimizing the maturity of the co-cultured muscle tissue. First, muscle tissues engineered from the C2C12 mouse myoblast cell line, cryopreserved primary human myoblasts, and freshly isolated primary chick myoblasts on micromolded gelatin hydrogels were compared. After three weeks, only chick muscle tissues remained stably adhered to hydrogels and exhibited progressive increases in myogenic index and stress generation, approaching values generated by native muscle tissue. After three weeks of co-culture with hiPSC-derived motor neurons, engineered chick muscle tissues formed NMJs with increasing co-localization of pre- and postsynaptic markers as well as increased frequency and magnitude of synaptic activity, surpassing structural and functional maturity of previous in vitro models. Engineered chick muscle tissues also demonstrated increased expression of genes related to sarcomere maturation and innervation over time, revealing new insights into the molecular pathways that likely contribute to enhanced NMJ formation. These approaches for engineering advanced neuromuscular tissues with relatively mature NMJs and interrogating their structure and function have many applications in neuromuscular disease modeling and drug development.
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影响因子：
3.8
作者：
Duffy RM;Sun Y;Feinberg AW
通讯作者：
Feinberg AW
影响因子：
2.7
作者：
DENNIS, MJ;ZISKINDCONHAIM, L;HARRIS, AJ
通讯作者：
HARRIS, AJ
影响因子：
16.6
作者：
Du, Zhong-Wei;Chen, Hong;Liu, Huisheng;Lu, Jianfeng;Qian, Kun;Huang, CindyTzu-Ling;Zhong, Xiaofen;Fan, Frank;Zhang, Su-Chun
通讯作者：
Zhang, Su-Chun
影响因子：
2.7
作者：
Biressi, Stefano;Bjornson, Christopher R. R.;Carlig, Poppy M. M.;Nishijo, Koichi;Keller, Charles;Rando, Thomas A.
通讯作者：
Rando, Thomas A.
影响因子：
3.7
作者：
Bloemberg D;Quadrilatero J
通讯作者：
Quadrilatero J