A comparative study of magnetic-activated cell sorting, cytotoxicity and preplating for the purification of human myoblasts.

Yonsei Medical Journal
Yoon Ghil ParkJin Kim

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.

References

Jan 1, 1988·Experimental Cell Research·C WebsterH M Blau
Dec 1, 1982·Journal of the Neurological Sciences·D J WattJ C Sloper
May 1, 1993·Muscle & Nerve·A BaroffioC R Bader
Sep 10, 1998·The Journal of Cell Biology·Z QuJ Huard
Jan 12, 1999·Muscle & Nerve·J R BlantonC A Bidwell

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Citations

Jul 21, 2009·Experimental Cell Research·Ulrich M BecherBernd K Fleischmann
Jun 11, 2015·Tissue Engineering. Part C, Methods·Shiplu Roy ChowdhuryRuszymah Bt Hj Idrus
Aug 23, 2008·Journal of Tissue Engineering and Regenerative Medicine·Mariea A BradyVivek Mudera
Jul 14, 2016·Tissue Engineering. Part C, Methods·Sushmitha DurgamMatthew Stewart
Sep 28, 2017·Tissue Engineering. Part C, Methods·Brian C SyverudLisa M Larkin
May 16, 2020·Drug Delivery and Translational Research·Nilesh RaiVibhav Gautam

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Methods Mentioned

BETA
fluorescence-activated
flow cytometry

Software Mentioned

SPSS

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