Lee Hyun, Han Na Rae, Kim Seong Jae, Yun Jung Im, Lee Seung Tae
Department of Animal Life Science, Kangwon National University, Chuncheon, Korea.
SCBIO Co., Ltd., Daejeon, Korea.
Int J Stem Cells. 2022 Aug 30;15(3):283-290. doi: 10.15283/ijsc21179. Epub 2022 Feb 28.
Difficulties often encountered in separating and purifying active muscle satellite cells (MSCs) from skeletal muscle tissues have limited the supply of cells for muscle therapy and artificial meat production. Here, we report an effective isolation protocol to economically and conveniently retrieve active MSCs from skeletal muscle tissues in mice.
We optimized an enzyme-based tissue digestion protocol for isolating skeletal muscle-derived primary cell population having a large number of active MSCs and described a method of differential plating (DP) for improving purity of active MSCs from skeletal muscle-derived primary cell population. Then, the age of the mouse appropriate to the isolation of a large number of active MSCs was elucidated. The best isolation yield of active MSCs from mouse skeletal muscle tissues was induced by the application of DP method to the primary cell population harvested from skeletal muscle tissues of 2-week-old mice digested in 0.2% (w/v) collagenase type II for 30 min at 37℃ and then in 0.1% (w/v) pronase for 5 min at 37℃.
The protocol we developed not only facilitates the isolation of MSCs but also maximizes the retrieval of active MSCs. Our expectation is that this protocol will contribute to the development of original technologies essential for muscle therapy and artificial meat industrialization in the future.
从骨骼肌组织中分离和纯化活性肌肉卫星细胞（MSCs）时经常遇到的困难，限制了用于肌肉治疗和人造肉生产的细胞供应。在此，我们报告一种有效的分离方案，可经济便捷地从小鼠骨骼肌组织中获取活性MSCs。
我们优化了一种基于酶的组织消化方案，用于分离具有大量活性MSCs的骨骼肌来源的原代细胞群体，并描述了一种差异铺板（DP）方法，以提高从骨骼肌来源的原代细胞群体中活性MSCs的纯度。然后，阐明了适合分离大量活性MSCs的小鼠年龄。通过将DP方法应用于从2周龄小鼠的骨骼肌组织中收获的原代细胞群体，该组织在37℃下用0.2%（w/v）II型胶原酶消化30分钟，然后在37℃下用0.1%（w/v）链霉蛋白酶消化5分钟，可诱导从小鼠骨骼肌组织中获得最佳的活性MSCs分离产量。
我们开发的方案不仅便于分离MSCs，而且能最大限度地获取活性MSCs。我们期望该方案将有助于未来肌肉治疗和人造肉产业化所需原创技术的发展。
