Optimization of culture of mesenchymal stem cells: a comparison of conventional plate and microcarrier cultures

Chang, Jing; Lv, Han; Liu, Q.; Qin, Shu; Ma, Ke; Luo, Shengjun; Zhang, X.; Huang, Wei; Zuo, Zhong; Fu, Hang; Xia, Yong

doi:10.1111/j.1365-2184.2012.00836.x

Cited by 18 publications

(13 citation statements)

References 15 publications

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“…Bioreactor‐facilitated stem cell expansion has already been achieved using multiple systems, ranging from multilayered cell factories to microcarriers in stirred tank bioreactors (Jung et al , ). Different groups have already shown that the stem cell phenotype can be maintained in these systems, despite the external hydrodynamic stresses applied to the cells by fluid flow during culture (Hewitt et al , ; Chang et al , ; Jung et al , ). However, despite the 3D shape of the carriers used in these systems, cells predominantly grow in a monolayer, which does not reproduce the native environment of the cells (Haycock, ).…”

Section: Introductionmentioning

confidence: 99%

Human periosteal-derived cell expansion in a perfusion bioreactor system: proliferation, differentiation and extracellular matrix formation

Sonnaert

Papantoniou

Bloemen

et al. 2014

J Tissue Eng Regen Med

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Perfusion bioreactor systems have shown to be a valuable tool for the in vitro development of three-dimensional (3D) cell-carrier constructs. Their use for cell expansion, however, has been much less explored. Since maintenance of the initial cell phenotype is essential in this process, it is imperative to obtain insight into the bioreactor-related variables determining cell fate. Therefore, this study investigated the influence of fluid flow-induced shear stress on the proliferation, differentiation and matrix deposition of human periosteal-derived cells in the absence of additional differentiation-inducing stimuli; 120 000 cells were seeded on additive manufactured 3D Ti6Al4V scaffolds and cultured for up to 28 days at different flow rates in the range 0.04-6 ml/min. DNA measurements showed, on average, a three-fold increase in cell content for all perfused conditions in comparison to static controls, whereas the magnitude of the flow rate did not have an influence. Contrast-enhanced nanofocus X-ray computed tomography showed substantial formation of an engineered neotissue in all perfused conditions, resulting in a filling (up to 70%) of the total internal void volume, and no flow rate-dependent differences were observed. The expression of key osteogenic markers, such as RunX2, OCN, OPN and Col1, did not show any significant changes in comparison to static controls after 28 days of culture, with the exception of OSX at high flow rates. We therefore concluded that, in the absence of additional osteogenic stimuli, the investigated perfusion conditions increased cell proliferation but did not significantly enhance osteogenic differentiation, thus allowing for this process to be used for cell expansion. Copyright © 2014 John Wiley & Sons, Ltd.

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Section: Introductionmentioning

confidence: 99%

Human periosteal-derived cell expansion in a perfusion bioreactor system: proliferation, differentiation and extracellular matrix formation

Sonnaert

Papantoniou

Bloemen

et al. 2014

J Tissue Eng Regen Med

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“…For MSCs, spreading shape versus round shape can change the fate of cell proliferation as well as osteogenic, adipogenic, or chondrogenic differentiation, which may be mediated through RhoA/ROCK signaling (Figure ). Despite the recent advances of MC technology for MSC cultivation, there are still significant challenges to fully exploit the potential of MCs for stem cell fate decision. There is clearly a limited understanding about how the MSCs attach to and interact with MCs.…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Engineering stem cell fate with biochemical and biomechanical properties of microcarriers

Sart

Agathos

2013

Biotechnology Progress

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Microcarriers have been widely used for various biotechnology applications because of their high scale-up potential, high reproducibility in regulating cellular behavior, and well-documented compliance with current Good Manufacturing Practices (cGMP). Recently, microcarriers have been emerging as a novel approach for stem cell expansion and differentiation, enabling potential scale-up of stem cell-derived products in large bioreactors. This review summarizes recent advances of using microcarriers in mesenchymal stem cell (MSC) and pluripotent stem cell (PSC) cultures. From the reported data, efficient expansion and differentiation of stem cells on microcarriers rely on their ability to modulate cell shape (i.e. round or spreading) and cell organization (i.e. aggregate size). Nonetheless, current screening of microcarriers remains empirical, and accurate understanding of how stem cells interact with microcarriers still remains unknown. This review suggests that accurate characterization of biochemical and biomechanical properties of microcarriers is required to fully exploit their potential in regulating stem cell fate decision. Due to the variety of microcarriers, such detailed analyses should lead to the rational design of application-specific microcarriers, enabling the exploitation of reproducible effects for large scale biomedical applications.

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“…This culture system showed some drawbacks such as cell aggregation during expansion. To avoid cell aggregation, some studies proposed controlling operating parameters such as agitation rate, cell density, and addition of carriers . Sometimes, due to the smooth surface of microcarriers such as glass microcarriers and Cytodex, MSCs cannot attach firmly and may detach and be washed away during medium changes .…”

Section: Introductionmentioning

confidence: 99%

Growth of human mesenchymal stem cells (MSCs) on films of enzymatically modified chitosan

Aljawish

Muniglia

Chevalot

2016

Biotechnology Progress

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in Wiley Online Library (wileyonlinelibrary.com)Mesenchymal stem cells (MSCs) are known to be an attractive cell source for tissue engineering and regenerative medicine. One of the main limiting steps for clinical use or biotechnological purposes is the expansion step. The research of compatible biomaterials for MSCs expansion is recently regarded as an attractive topic. The aim of this study was to create new functional biomaterial for MSCs expansion by evaluating the impact of chitosan derivative films modified by enzymatic approach. First, chitosan particles were enzymatically modified with ferulic acid (FA) or ethyl ferulate (EF) under an eco-friendly procedure. Then, films of chitosan and its modified derivatives were prepared and evaluated by physicochemical and biological properties. Results showed that the enzymatic grafting of FA or EF onto chitosan significantly increased hydrophobic and antioxidant properties of chitosan films. The MSCs cell viability on chitosan derivative films also increased depending on the film thickness and the quantity of grafted phenols. Furthermore, the cytotoxicity test showed the absence of toxic effect of chitosan derivative films towards MSCs cells. Cell morphology showed a well attached and spread phenotype of MSCs cells on chitosan derivative films. On the other hand, due to the higher phenol content of FA-chitosan films, their hydrophobic, antioxidant properties and cell adhesion were improved in comparison with those of EFchitosan films. Finally, this enzymatic process can be considered as a promising process to favor MSCs cell growth as well as to create useful biomaterials for biomedical applications especially for tissue engineering.

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Optimization of culture of mesenchymal stem cells: a comparison of conventional plate and microcarrier cultures

Abstract: Compared to conventional plate culture, culture using gelatine porous microcarrier Cultispher-S was superior for large-scale production of rat MSCs.

Cited by 18 publications

References 15 publications

Human periosteal-derived cell expansion in a perfusion bioreactor system: proliferation, differentiation and extracellular matrix formation

Human periosteal-derived cell expansion in a perfusion bioreactor system: proliferation, differentiation and extracellular matrix formation

Engineering stem cell fate with biochemical and biomechanical properties of microcarriers

Growth of human mesenchymal stem cells (MSCs) on films of enzymatically modified chitosan

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