A Comparative Study of Magnetic-Activated Cell Sorting, Cytotoxicity and Preplating for the Purification of Human Myoblasts

Park, Yoon Ghil; Moon, Joon-Ho; Kim, Jin

doi:10.3349/ymj.2006.47.2.179

Cited by 25 publications

(16 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To date, a number of researchers have reported several myoblast purification procedures to obtain a homogeneous myogenic cell culture, including density centrifugation, selective adhesion and chemical use of fibroblast growth inhibitors (Kao and Prockop, 1977;Yablonka-Reuveni et al, 1987;Blanton et al, 1999). Immunomagnetic techniques have recently been reported (Lequerica et al, 1999;Sinanan et al, 2004;Park et al, 2006) to purify myogenic cells from primary cultures by magnetic activated cell sorting, with good yields. Lequerica et al (1999) reported an increase in the percentage of myoblasts in mixed culture from 8.4% to more than 90% using this technique.…”

Section: Discussionmentioning

confidence: 99%

Synergy between myogenic and non-myogenic cells in a 3D tissue-engineered craniofacial skeletal muscle construct

Brady

Lewis

Mudera

2008

J Tissue Eng Regen Med

View full text Add to dashboard Cite

In vitro skeletal muscle engineering involves the culture of isolated primary myogenic cells in an environment conducive to the formation of a three-dimensional (3D) tissue construct capable of generating force. Isolated human myogenic cells have been used to study cell-cell interactions, permitting identification of functions intrinsic to skeletal muscle in two dimensions (2D). However, the independent contribution of human myogenic and non-myogenic cell types that comprise skeletal muscle to myogenic cell differentiation, force generation and matrix remodelling has yet to be established in 3D. The objective of this study was to use isolated human myogenic and non-myogenic muscle-derived cells (MDC) seeded in 3D collagen constructs to engineer a biomimetic craniofacial skeletal construct. The aim was to purify the two subpopulations of myogenic and non-myogenic cells from human masseter muscle and quantitate myogenic cell differentiation, force generation and matrix remodelling of the 3D collagen construct. The results showed that both the heterogeneous mixture of cells and the purified myogenic cell population expressed myogenin, indicative of myogenic cell differentiation. Further, there was a synergistic effect as the heterogeneous co-culture of myogenic and non-myogenic cells generated the highest peak force and expressed the most MMP-2 mRNA compared to isolated individual cell populations.

show abstract

Section: Discussionmentioning

confidence: 99%

Synergy between myogenic and non-myogenic cells in a 3D tissue-engineered craniofacial skeletal muscle construct

Brady

Lewis

Mudera

2008

J Tissue Eng Regen Med

View full text Add to dashboard Cite

show abstract

“…Primary skeletal muscle stem cells were isolated from a M. vastus lateralis muscle biopsy as described (Orth et al, 2003;Park et al, 2006) (Fig. 1c).…”

Section: Generation Of Myoblast From Human Muscle Biopsiesmentioning

confidence: 99%

“…Cells were split using TrypLE (Life technologies). To enrich myogenic cells the pre-plating method described by Park and collaborators (Park et al, 2006) with some modifications was used. Briefly, cells were detached and then pre-plated in a gelatincoated flask.…”

Section: Differentiation Of Hipsc Into Myoblasts or Motoneuronsmentioning

confidence: 99%

Formation and characterisation of neuromuscular junctions between hiPSC derived motoneurons and myotubes

Demestre

Orth

Föhr³

et al. 2015

Stem Cell Research

View full text Add to dashboard Cite

Striated skeletal muscle cells from humans represent a valuable source for in vitro studies of the motoric system as well as for pathophysiological investigations in the clinical settings. Myoblasts can readily be grown from human muscle tissue. However, if muscle tissue is unavailable, myogenic cells can be generated from human induced pluripotent stem cells (hiPSCs) preferably without genetic engineering. Our study aimed to optimize the generation of hiPSCs derived myogenic cells by employing selection of CD34 positive cells and followed by distinct, stepwise culture conditions. Following the expansion of CD34 positive single cells under myogenic cell culture conditions, serum deprived myoblast-like cells finally fused and formed multinucleated striated myotubes that expressed a set of key markers for muscle differentiation. In addition, these myotubes contracted upon electrical stimulation, responded to acetylcholine (Ach) and were able to generate action potentials. Finally, we co-cultured motoneurons and myotubes generated from identical hiPSCs cell lines. We could observe the early aggregation of acetylcholine receptors in muscle cells of immature co-cultures. At later stages, we identified and characterised mature neuromuscular junctions (NMJs). In summary, we describe here the successful generation of an iPS cell derived functional cellular system consisting of two distinct communicating cells types. This in vitro co-culture system could therefore contribute to research on diseases in which the motoneurons and the NMJ are predominantly affected, such as in amyotrophic lateral sclerosis or spinal muscular atrophy.

show abstract

“…For tissue engineers, knowing that final purity is essential for consistent tissue fabrication. Fluorescence activated cell sorting (FACS) and magnetic activated cell sorting (MACS) have been used to evaluate the success of the pre-plating process, leading researchers to consider whether they can be used to skip pre-plating and immediately sort instead [51,52]. …”

Section: Purificationmentioning

confidence: 99%

“…Research by the Blau laboratory demonstrated that semi-pure mouse myoblast populations may be isolated using MACS technology to select for α7 Integrin + cells, with similar purification levels as found with FACS [52]. Although MACS has been used in some cases to purify CD56 positive satellite cells [62], direct comparison of pre-plating and MACS indicated much greater purification using the pre-plating technique [51]. From these studies, it is clear that multiple markers are likely needed isolate a highly pure population.…”

Section: Purificationmentioning

confidence: 99%

Isolation and Purification of Satellite Cells for Skeletal Muscle Tissue Engineering

et al. 2015

View full text Add to dashboard Cite

Engineered skeletal muscle holds promise as a source of graft tissue for the repair of traumatic injuries such as volumetric muscle loss. The resident skeletal muscle stem cell, the satellite cell, has been identified as an ideal progenitor for tissue engineering due to its role as an essential player in the potent skeletal muscle regeneration mechanism. A significant challenge facing tissue engineers, however, is the isolation of sufficiently large satellite cell populations with high purity. The two common isolation techniques, single fiber explant culture and enzymatic dissociation, can yield either a highly pure satellite cell population or a suitably large number or cells but fail to do both simultaneously. As a result, it is often necessary to use a purification technique such as pre-plating or cell sorting to enrich the satellite cell population post-isolation. Furthermore, the absence of complex chemical and biophysical cues influencing the in vivo satellite cell “niche” complicates the culture of isolated satellite cells. Techniques under investigation to maximize myogenic proliferation and differentiation in vitro are described in this article, along with current methods for isolating and purifying satellite cells.

show abstract

A Comparative Study of Magnetic-Activated Cell Sorting, Cytotoxicity and Preplating for the Purification of Human Myoblasts

Cited by 25 publications

References 23 publications

Synergy between myogenic and non-myogenic cells in a 3D tissue-engineered craniofacial skeletal muscle construct

Synergy between myogenic and non-myogenic cells in a 3D tissue-engineered craniofacial skeletal muscle construct

Formation and characterisation of neuromuscular junctions between hiPSC derived motoneurons and myotubes

Isolation and Purification of Satellite Cells for Skeletal Muscle Tissue Engineering

Contact Info

Product

Resources

About