A Comparative Study of Magnetic-Activated Cell Sorting, Cytotoxicity and Preplating for the Purification of Human Myoblasts.
10.3349/ymj.2006.47.2.179
- Author:
Yoon Ghil PARK
1
;
Jae Ho MOON
;
Jin KIM
Author Information
1. Department of Rehabilitation Medicine and Rehabilitation Institute of Muscular Disease, Yonsei University College of Medicine, Seoul, Korea. drtlc@yumc.yonsei.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Muscular dystrophy;
myoblast;
magnetic-activated cell sorting;
complement-mediated cytotoxicity;
preplating
- MeSH:
Time Factors;
Myoblasts/*cytology;
Muscle, Skeletal/*cytology;
Models, Statistical;
Magnetics;
Immunomagnetic Separation/methods;
Immunohistochemistry;
Humans;
Genetic Complementation Test;
Fibroblasts/cytology;
Complement System Proteins;
Cells, Cultured;
Cell Separation/*methods;
Cell Differentiation
- From:Yonsei Medical Journal
2006;47(2):179-183
- CountryRepublic of Korea
- Language:English
-
Abstract:
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.
