Pax3-induced expansion enables the genetic correction of dystrophic satellite cells.
Pax3-induced expansion enables the genetic correction of dystrophic satellite cells.
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DOI：
10.1186/s13395-015-0061-7
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发表时间：
2015
期刊：
影响因子：
4.9
通讯作者：
Perlingeiro RC
中科院分区：
文献类型：
--
作者：
Filareto A;Rinaldi F;Arpke RW;Darabi R;Belanto JJ;Toso EA;Miller AZ;Ervasti JM;McIvor RS;Kyba M;Perlingeiro RC
卫星细胞（SC）对于肌肉再生和修复不可或缺；然而，由于初级肌肉中的频率低以及离体扩增后植入潜力的丧失，它们在细胞治疗中的使用目前是不可行的。迄今为止，解决这一限制的替代方案是移植 SC 衍生的肌源祖细胞 (MPC)，尽管它们不具有干细胞相同的有吸引力的特性，例如自我更新和长期再生潜力。我们开发了一种使用 Pax3 瞬时表达来扩增野生型和营养不良的新鲜分离卫星细胞的方法。这种方法可以与营养不良卫星细胞的遗传校正相结合，并在移植到营养不良小鼠体内时用于促进肌肉再生。在这里，我们证明来自野生型和营养不良小鼠的 SC 可以通过 Pax3 的瞬时表达在培养物中扩增，并且这些扩增的活化 SC 可以再生肌肉。我们在基因治疗模型中测试了这种方法，通过使用携带人类 μDYSTROPHIN 基因的睡美人转座子来纠正缺乏肌营养不良蛋白的小鼠的营养不良 SC。将这些扩增的校正细胞移植到免疫缺陷、肌营养不良蛋白缺陷的小鼠中，产生了大量表达肌营养不良蛋白的肌纤维并改善了收缩强度。重要的是，体外扩增的 SC 移植到 SC 隔室中，并且可以在二次损伤后再生肌肉。这些结果表明 Pax3 能够促进 SC 的离体扩增，同时保持其干细胞再生特性。本文的在线版本 (doi:10.1186/s13395-015-0061-7) 包含补充材料，可供授权用户使用。
Satellite cells (SCs) are indispensable for muscle regeneration and repair; however, due to low frequency in primary muscle and loss of engraftment potential after ex vivo expansion, their use in cell therapy is currently unfeasible. To date, an alternative to this limitation has been the transplantation of SC-derived myogenic progenitor cells (MPCs), although these do not hold the same attractive properties of stem cells, such as self-renewal and long-term regenerative potential. We develop a method to expand wild-type and dystrophic fresh isolated satellite cells using transient expression of Pax3. This approach can be combined with genetic correction of dystrophic satellite cells and utilized to promote muscle regeneration when transplanted into dystrophic mice. Here, we show that SCs from wild-type and dystrophic mice can be expanded in culture through transient expression of Pax3, and these expanded activated SCs can regenerate the muscle. We test this approach in a gene therapy model by correcting dystrophic SCs from a mouse lacking dystrophin using a Sleeping Beauty transposon carrying the human μDYSTROPHIN gene. Transplantation of these expanded corrected cells into immune-deficient, dystrophin-deficient mice generated large numbers of dystrophin-expressing myofibers and improved contractile strength. Importantly, in vitro expanded SCs engrafted the SC compartment and could regenerate muscle after secondary injury. These results demonstrate that Pax3 is able to promote the ex vivo expansion of SCs while maintaining their stem cell regenerative properties. The online version of this article (doi:10.1186/s13395-015-0061-7) contains supplementary material, which is available to authorized users.