Minimally Invasive Muscle Embedding (MIME) - A Novel Experimental Technique to Facilitate Donor-Cell-Mediated Myogenesis.
Minimally Invasive Muscle Embedding (MIME) - A Novel Experimental Technique to Facilitate Donor-Cell-Mediated Myogenesis.
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DOI：
10.3791/55731
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发表时间：
2017-08-24
期刊：
影响因子：
--
通讯作者：
Galen SS
中科院分区：
文献类型：
--
作者：
Roche JA;Begam M;Galen SS
骨骼肌具有再生能力，这是由于组织驻留的肌纤维生成（肌原性）卫星细胞（SC），可以在适当的条件下形成新的肌纤维。虽然可以从肌肉组织中收获SC并在体外培养，但当移植到宿主肌肉中时，所得成肌细胞在促进肌生成方面不是非常有效。与成肌细胞移植相比，手术暴露宿主肌肉并移植供体肌肉组织的片段，或分离的肌纤维及其SC到宿主肌肉上，促进更好的肌发生。我们开发了一种新技术，我们称之为微创肌肉嵌入（MIME）。MIME涉及将手术针穿过宿主肌肉，通过针道吸引一块供体肌肉组织，然后将供体组织嵌入宿主肌肉中，以便其可以充当宿主肌肉的SC来源。在这里，我们详细描述了在免疫缺陷小鼠模型中进行MIME所涉及的步骤，该模型在其所有细胞中表达绿色荧光蛋白（GFP）。宿主小鼠中的免疫缺陷降低了供体组织的免疫排斥的风险，并且GFP表达使得能够容易地鉴定宿主肌纤维（GFP+）和供体细胞衍生的肌纤维（GFP-）。我们的试验数据表明，MIME可以用于植入伸趾长肌（EDL）肌肉从供体小鼠到胫骨前（TA）肌肉的宿主小鼠。我们的数据还表明，当肌肉毒素（氯化钡，氯化钡）注射到主机肌肉后MIME，有证据表明供体细胞来源的肌发生在主机肌肉，约5%，26%，26%和43%的纤维在一个单一的主机TA肌肉显示没有主机的贡献，最小的主机贡献，中等主机的贡献，和最大的主机贡献，分别。
Skeletal muscle possesses regenerative capacity due to tissue-resident, muscle-fiber-generating (myogenic) satellite cells (SCs), which can form new muscle fibers under the right conditions. Although SCs can be harvested from muscle tissue and cultured in vitro, the resulting myoblast cells are not very effective in promoting myogenesis when transplanted into host muscle. Surgically exposing the host muscle and grafting segments of donor muscle tissue, or the isolated muscle fibers with their SCs onto host muscle, promotes better myogenesis compared to myoblast transplantation. We have developed a novel technique that we call Minimally Invasive Muscle Embedding (MIME). MIME involves passing a surgical needle through the host muscle, drawing a piece of donor muscle tissue through the needle track, and then leaving the donor tissue embedded in the host muscle so that it may act as a source of SCs for the host muscle. Here we describe in detail the steps involved in performing MIME in an immunodeficient mouse model that expresses a green fluorescent protein (GFP) in all of its cells. Immunodeficiency in the host mouse reduces the risk of immune rejection of the donor tissue, and GFP expression enables easy identification of the host muscle fibers (GFP+) and donor-cell-derived muscle fibers (GFP-). Our pilot data suggest that MIME can be used to implant an extensor digitorum longus (EDL) muscle from a donor mouse into the tibialis anterior (TA) muscle of a host mouse. Our data also suggest that when a myotoxin (barium chloride, BaCl2) is injected into the host muscle after MIME, there is evidence of donor-cell-derived myogenesis in the host muscle, with approximately 5%, 26%, 26% and 43% of the fibers in a single host TA muscle showing no host contribution, minimal host contribution, moderate host contribution, and maximal host contribution, respectively.
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影响因子：
4.9
作者：
Sakellariou P;O'Neill A;Mueller AL;Stadler G;Wright WE;Roche JA;Bloch RJ
通讯作者：
Bloch RJ
影响因子：
4.1
作者：
Lin, Hang;Cheng, Anthony Wai-Ming;Tuan, Rocky S.
通讯作者：
Tuan, Rocky S.
影响因子：
4.9
作者：
Krag, Thomas O.;Hauerslev, Simon;Vissing, John
通讯作者：
Vissing, John
影响因子：
12.4
作者：
Skuk, D;Roy, B;Tremblay, JP
通讯作者：
Tremblay, JP
影响因子：
3.7
作者：
Hardy D;Besnard A;Latil M;Jouvion G;Briand D;Thépenier C;Pascal Q;Guguin A;Gayraud-Morel B;Cavaillon JM;Tajbakhsh S;Rocheteau P;Chrétien F
通讯作者：
Chrétien F