A comparative study of magnetic-activated cell sorting, cytotoxicity and preplating for the purification of human myoblasts.
Although cultured myoblast transplantation has been extensively studied as a gene complementation approach to muscular dystrophy treatment, clinical success has still been limited. The inability to adequately isolate and purify myoblasts presents a major limitation to the production of sufficient myoblasts for engrafting purposes. This study attempted to purify myoblasts from primary culture by magnetic-activated cell sorting (MACS), complement-mediated cytotoxicity, and a preplating technique. As a result of positive myoblasts selection by MACS, the average percentage of myoblasts in mixed culture was increased from 30.0% to 41.7%. We observed both myoblast lysis and fibroblast lysis after complement-mediated cytotoxicity. Enrichment of myoblasts in mixed culture was found to increase to 83.1% by using the preplating technique. In addition, higher purification (92.8%) was achieved by following the preplating technique with MACS. Thus, preplating in combination with magnetic-activated cell sorting allows for a rapid and effective isolation of myoblasts from human muscle tissue.
Park
Yoon Ghil
YG
Department of Rehabilitation Medicine, Yongdong Severance Hospital, Yonsei University College, 146-92 Dogok-dong, Gangnam-gu, Seoul 135-270, Korea. drtlc@yumc.yonsei.ac.kr
Moon
Jae Ho
JH
Kim
Jin
J
D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D003165:Complement System Proteins; D005347:Fibroblasts / Q000166:cytology; D005816:Genetic Complementation Test; D006801:Humans; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation / Q000379:methods; D008280:Magnetics; D015233:Models, Statistical; D018482:Muscle, Skeletal / Q000166:cytology*; D032446:Myoblasts / Q000166:cytology*; D013997:Time Factors
D003165:Complement System Proteins