Thermoresponsive Substrates used for the Expansion of Human Mesenchymal Stem Cells and the Preservation of Immunophenotype

Nash, Maria E.; Fan, Xingliang; Carroll, William M.; Gorelov, Alexander; Barry, Frank; Shaw, Georgina; Rochev, Yury

doi:10.1007/s12015-013-9428-5

Cited by 29 publications

(31 citation statements)

References 28 publications

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“…). The reason why the pNIPAm films become increasingly cell repulsive with increasing film thickness is unclear though it is probably associated with the increased mobility of the polymer chains with increasing film thickness leading to increased hydration which is non‐conducive to protein adsorption which mediates cell attachment, this increased hydrophilicity for the thicker 4 µm thick films was analytically reflected in the advancing contact angle measurements …”

Section: Resultsmentioning

confidence: 99%

“…For solvent cast films, 2% w/v polymer solution in EtOH was spread on 35 mm TCP. The thickness of the solvent cast film can be estimated from total mass of polymer spread over defined area of Petri dish assuming dry polymer density 1 g cm –3 …”

Section: Methodsmentioning

confidence: 99%

“…The film was used for the multifold passaging of hMSCs. It was found that cell proliferation through the passages was similar in all cases and the pluripotent nature of the hMSCs was preserved . Another example is the use of thermoresponsive nanocomposite gels to proliferate and harvest human mesenchymal stem cells (hMSCs), which should be effectively cultured in vitro for tissue engineering and regenerative medicine .…”

Section: Introductionmentioning

confidence: 99%

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Macrophages behavior on different NIPAm‐based thermoresponsive substrates

Fan

Nosov

Carroll

et al. 2013

J Biomedical Materials Res

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Thermoresponsive materials and surfaces are widely used in cell culture applications. There is a lot of research work employing thermoresponsive materials with various structure and compositions. However, little is known about the immunological response to the thermoresponsive materials. Macrophage‐like transformed murine cell line RAW264.7 was selected as it is a widely used standard model for immune activation analysis. This study proposes to compare the effects of thermoresponsive films with various compositions on macrophage cells. Thermoresponsive materials are a useful utility as a non‐enzymatic harvesting system for tissue culture. As RAW264.7 cells are difficult to remove from the substrate by enzymatic methods we also explored the possibility to use thermoresponsive materials for the macrophage cultivation. Spin coating and solvent casting was used to produce films of N‐isopropylacrylamide‐based polymers from the nanometer to micrometer range. Successful cell adhesion and proliferation was highly dependent on the thickness and composition of the coating. RAW264.7 cells were successfully detached from the coatings upon temperature reduction. Furthermore, results indicate that the RAW264.7 cells remained inactivated as cell secreted cytokine remained at a low level and the surface receptor profile of RAW264.7 was not altered when cells were detached in this manner. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 2901–2910, 2014.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Macrophages behavior on different NIPAm‐based thermoresponsive substrates

Fan

Nosov

Carroll

et al. 2013

J Biomedical Materials Res

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“…We used the seeding density of 8000 cells/cm 2 as a control in this experiment, since this seeding density is often used for expansion [7]. The seeding densities 20,000, 50,000 and 400,000 cells/cm 2 were selected based on previous studies examining cell sheets [10, 26, 36]. After overnight attachment, we refreshed the cells with LG-DMEM with 1% FBS, 50 μg/ml gentamicin and 1.5 μg/ml fungizone® (control condition).…”

Section: Methodsmentioning

confidence: 99%

Effect of Cell Seeding Density and Inflammatory Cytokines on Adipose Tissue-Derived Stem Cells: an in Vitro Study

Sukho

Kirpensteijn

Hesselink

et al. 2017

Stem Cell Rev and Rep

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Adipose tissue-derived stem cells (ASCs) are known to be able to promote repair of injured tissue via paracrine factors. However, the effect of cell density and inflammatory cytokines on the paracrine ability of ASCs remains largely unknown. To investigate these effects, ASCs were cultured in 8000 cells/cm2, 20,000 cells/cm2, 50,000 cells/cm2, and 400,000 cells/cm2 with and without 10 or 20 ng/ml tumor necrosis factor alpha (TNFα) and 25 or 50 ng/ml interferon gamma (IFNγ). ASC-sheets formed at 400,000 cells/cm2 after 48 h of culture. With increasing concentrations of TNFα and IFNγ, ASC-sheets with 400,000 cells/cm2 had increased production of angiogenic factors Vascular Endothelial Growth Factor and Fibroblast Growth Factor and decreased expression of pro-inflammatory genes TNFA and Prostaglandin Synthase 2 (PTGS2) compared to lower density ASCs. Moreover, the conditioned medium of ASC-sheets with 400,000 cells/cm2 stimulated with the low concentration of TNFα and IFNγ enhanced endothelial cell proliferation and fibroblast migration. These results suggest that a high cell density enhances ASC paracrine function might beneficial for wound repair, especially in pro-inflammatory conditions.Electronic supplementary materialThe online version of this article (doi:10.1007/s12015-017-9719-3) contains supplementary material, which is available to authorized users.

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“…To date, PNIPAm has been the most employed synthetic polymer to prepare thermo-responsive MIPs, due to its LCST which is close to the temperature of the human body [93]. One of the earliest thermo-responsive MIPs was reported by Watanabe et al [87].…”

Section: Thermo-responsive Molecularly Imprinted Polymersmentioning

confidence: 99%

Molecularly Imprinted Polymers with Stimuli-Responsive Affinity: Progress and Perspectives

et al. 2015

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Intelligent stimuli-responsive molecularly imprinted polymers (SR-MIPs) have attracted considerable research interest in recent years due to the potential applications in drug delivery, biotechnology and separation sciences. This review comprehensively summarizes various SR-MIPs, including the design and applications of thermo-responsive MIPs, pH-responsive MIPs, photo-responsive MIPs, biomolecule-responsive MIPs and ion-responsive MIPs. Besides the development of current SR-MIPs, the advantages as well as the disadvantages of current SR-MIPs were also displayed from different angles, especially preparation methods and application fields. We believe this review will be helpful to guide the design, development and application of SR-MIPs.Polymers 2015, 7 1690

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Thermoresponsive Substrates used for the Expansion of Human Mesenchymal Stem Cells and the Preservation of Immunophenotype

Cited by 29 publications

References 28 publications

Macrophages behavior on different NIPAm‐based thermoresponsive substrates

Macrophages behavior on different NIPAm‐based thermoresponsive substrates

Effect of Cell Seeding Density and Inflammatory Cytokines on Adipose Tissue-Derived Stem Cells: an in Vitro Study

Molecularly Imprinted Polymers with Stimuli-Responsive Affinity: Progress and Perspectives

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