Maintenance of muscle fiber-type memory in a human-to-mouse muscle xenograft model

Abstract

INTRODUCTION. Muscle tissue can regenerate through muscle stem cells known as muscle satellite cells (MuSCs). MIME is a technique developed in our laboratory, through which, donor muscle tissue with MuSCs can be implanted into a host muscle to promote donor-cell-derived myogenesis. OBJECTIVE. We studied whether or not MuSCs maintain memory of the type of muscle fibers (fast or slow) with which they were associated. We hypothesized that MuSCs do not maintain memory of the fibers with which they are associated and form slow or fast muscle fibers based on the host environment. METHODS. We tested our hypothesis by implanting donor muscle, which contained mostly slow fibers, into host muscle, which contained mostly fast fibers. Implantation of donor tissue was performed by MIME. Following MIME, we injected a myotoxin (1.2% barium chloride, BaCl2) into the host muscle, which induced concerted degeneration followed by MuSC-dependent regeneration. Specifically, we studied mouse tibialis anterior muscles, which were implanted with segments of human donor tibialis anterior muscle. The human TA is comprised mostly of slow fibers (~60%; i.e., slow myosin heavy chain positive [sMyHC+]). The mouse TA is comprised 100% of fast fibers (i.e., fast myosin heavy chain positive [fMyHC+] fibers). This experimental paradigm is appropriate to ascertain whether or not satellite cells retain memory of the muscle fibers with which they were originally associated. We collected host muscles after 12 weeks of regeneration to ascertain if there was an increase in the percentage of slow muscle fibers. We used antibodies specific to sMyHC and fMyHC to stain frozen TA muscle sections. Host mice were immunodeficient and ubiquitously expressed enhanced green fluorescent protein (NSG-EGFP, The Jackson Laboratory Strain #021937). Since host mouse muscle fibers exhibited green fluorescence, but donor human muscle fibers did not, host and donor muscle fibers could be easily differentiated under fluorescence optics. RESULTS. Quantitative data confirmed that the cluster of human muscle fibers, which regenerated in the host mouse TA muscle had more slow (~50%) than fast (~30%) fibers (T-test, p<0.01, N = 4 TA muscles from 4 different host mice). CONCLUSION. Contrary to our hypothesis, our data suggest that MuSCs do maintain memory of the type of muscle fibers with which they were originally associated. MuSCs give rise to fast or slow muscle fibers based on their donor origin when they are implanted along with their original donor muscle tissue. This work was supported by NIH R03HD091648, Pitch Award from the Alliance for Regenerative Rehabilitation Research and Training (AR3T) from NIH P2CHD086843, and a subcontract from NIH R01AR079884-01 (Peter L. Jones PI) to JAR. We thank the Wayne State University DPT Program for supporting student research and providing research training through coursework and research experiences. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.