Cell Shape and Cardiosphere Differentiation: A Revelation by Proteomic Profiling

Stem cells have emerged as promising tools for the treatment of incurable neural and heart diseases and tissue damage. However, the survival of transplanted stem cells is reported to be low, reducing their therapeutic effects. The major causes of poor survival of stem cells in vivo are linked to anoikis, potential immune rejection, and oxidative damage mediating apoptosis. This review investigates novel methods and potential molecular mechanisms for stem cell preconditioning in vitro to increase their retention after transplantation in damaged tissues. Microenvironmental preconditioning (e.g., hypoxia, heat shock, and exposure to oxidative stress), aggregate formation, and hydrogel encapsulation have been revealed as promising strategies to reduce cell apoptosis in vivo while maintaining biological functions of the cells. Moreover, this review seeks to identify methods of optimizing cell dose preparation to enhance stem cell survival and therapeutic function after transplantation.

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“… 74 Similarly, cardiospheres derived from CPCs showed enhanced glycolysis, possibly as a consequence of the formation of hypoxic area. 75 …”

Section: Preconditioning Stem Cells As Aggregatesmentioning

confidence: 99%

Preconditioning Stem Cells for In Vivo Delivery

Sart

2014

BioResearch Open Access

160

142

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“…Stem cells are known to form spherical (i.e., 3D) embryonic bodies (EBs). To compare 2D and 3D states, we recently compared the proteomic profiles of sphere-forming and adherent cells using CSC-21E [34, 47]. In this experiment, the same culture medium was used; however the dish substrate was changed.…”

Section: Proteome Analysismentioning

confidence: 99%

3D-Culture System for Heart Regeneration and Cardiac Medicine

Kawaguchi

Hatta

Nakanishi

2013

BioMed Research International

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3D cultures have gained attention in the field of regenerative medicine for their usefulness as in vitro model of solid tissues. Bottom-up technology to generate artificial tissues or organs is prospective and an attractive approach that will expand as the field of regenerative medicine becomes more translational. We have characterized c-kit positive cardiac stem cells after long-term cultures and established a 3D-nanoculture system using collagen scaffolds. By combining informatics-based studies, including proteomic analyses and microarrays, we sought to generate methods that modeled cardiac regeneration which can ultimately be used to build artificial hearts. Here, we describe the use of biodegradable beads or 3D cultures to study cardiac regeneration. We summarize recent work that demonstrates that, by using a combination of molecular analyses with 3D cultures, it is possible to evaluate concise mechanisms of solid tissue stem cell biology.

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“…Accordingly, the reliability of the method used to estimate these parameters plays a key role in this analysis [ 8 ]. In addition, cell counting and viability assessment often need to be performed for other 3D cell cultures, such as stem cell spheroids generated for regenerative medicine purposes [ 9 ], and organoids used to study (some) organ characteristics [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Cell Counting and Viability Assessment of 2D and 3D Cell Cultures: Expected Reliability of the Trypan Blue Assay

et al. 2017

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BackgroundWhatever the target of an experiment in cell biology, cell counting and viability assessment are always computed. The Trypan Blue (TB) assay was proposed about a century ago and is still the most widely used method to perform cell viability analysis. Furthermore, the combined use of TB with a haemocytometer is also considered the standard approach to estimate cell population density. There are numerous research articles reporting the use of TB assays to compute cell number and viability of 2D and 3D cultures. However, the literature still lacks studies regarding the reliability of the TB assay in terms of assessment of its repeatability and reproducibility.MethodsWe compared the TB assay's measurements obtained by two biologists who analysed 105 different samples in double-blind for a total of 210 counts performed. We measured: (a) the repeatability of the count performed by the same operator; (b) the reproducibility of counts performed by the two operators.ResultsThere were no significant differences in the results obtained with 2D and 3D cell cultures: we estimated an approximate variability of 5% when the TB assay was used to assess the viability of the culture, and a variability of around 20% when it was used to determine the cell population density.ConclusionsThe main aim of this study was to make researchers aware of potential measurement errors when TB is used with a haemocytometer for counting and viability measurements in 2D and 3D cultures. We believe that these results can help researchers to determine whether the expected reliability of the TB assay is compliant with their applications.

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Cell Shape and Cardiosphere Differentiation: A Revelation by Proteomic Profiling

Cited by 9 publications

References 93 publications

Preconditioning Stem Cells for In Vivo Delivery

Preconditioning Stem Cells for In Vivo Delivery

3D-Culture System for Heart Regeneration and Cardiac Medicine

Cell Counting and Viability Assessment of 2D and 3D Cell Cultures: Expected Reliability of the Trypan Blue Assay

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