Walthers Christopher M, Lee Min, Wu Benjamin M, Dunn James C Y
Department of Bioengineering, University of California Los Angeles, Los Angeles, California, United States of America.
Division of Advanced Prosthodontics, Biomaterials, and Hospital Dentistry, University of California Los Angeles, Los Angeles, California, United States of America; The Weintraub Center for Reconstructive Biotechnology, University of California Los Angeles, Los Angeles, California, United States of America.
PLoS One. 2014 Dec 8;9(12):e114850. doi: 10.1371/journal.pone.0114850. eCollection 2014.
Functionally contracting smooth muscle is an essential part of the engineered intestine that has not been replicated in vitro. The purpose of this study is to produce contracting smooth muscle in culture by maintaining the native smooth muscle organization. We employed intact smooth muscle strips and compared them to dissociated smooth muscle cells in culture for 14 days. Cells isolated by enzymatic digestion quickly lost maturity markers for smooth muscle cells and contained few enteric neural and glial cells. Cultured smooth muscle strips exhibited periodic contraction and maintained neural and glial markers. Smooth muscle strips cultured for 14 days also exhibited regular fluctuation of intracellular calcium, whereas cultured smooth muscle cells did not. After implantation in omentum for 14 days on polycaprolactone scaffolds, smooth muscle strip constructs expressed high levels of smooth muscle maturity markers as well as enteric neural and glial cells. Intact smooth muscle strips may be a useful component for engineered intestinal smooth muscle.
功能性收缩的平滑肌是工程化肠道的重要组成部分，尚未在体外复制。本研究的目的是通过维持天然平滑肌组织在培养物中产生收缩性平滑肌。我们使用完整的平滑肌条，并将它们与培养14天的解离平滑肌细胞进行比较。通过酶消化分离的细胞迅速失去平滑肌细胞的成熟标志物，并且含有很少的肠神经和神经胶质细胞。培养的平滑肌条表现出周期性收缩，并维持神经和神经胶质标志物。培养14天的平滑肌条也表现出细胞内钙的规律性波动，而培养的平滑肌细胞则没有。在聚己内酯支架上植入大网膜14天后，平滑肌条构建体表达高水平的平滑肌成熟标志物以及肠神经和神经胶质细胞。完整的平滑肌条可能是工程化肠道平滑肌的有用组成部分。
