Bulk poly(<i>N</i>-isopropylacrylamide) (PNIPAAm) thermoresponsive cell culture platform: toward a new horizon in cell sheet engineering

Choi, Andrew; Seo, Kyoung Duck; Yoon, Hyungjun; Han, Seon Jin; Kim, Dong Sung

doi:10.1039/c8bm01664j

Cited by 41 publications

(28 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…PNIPAAm has been used for smart materials, which mainly utilizes surface property changes and volume phase transitions with response to ambient temperature. The versatility of PNIPAAm has led to finding uses in cell culture [4], tissue engineering [5], enzymatic immobilization [6], drug delivery [7], wound dressing [8], biosensors [9], etc. One of the most important applications of PNIPAAm is cell culture.…”

Section: Preparation Of Pnipaam Medicated Substrates For Cell Culturementioning

confidence: 99%

“…The permeable calcein-AM is enzymatically converted into calcein showing bright green fluorescence in the cytoplasm of viable cells, while ethidium homodimer can enter cells with compromised membranes and takes up red fluorescence upon binding to nucleic acids [114]. The ratio of fluorescence intensity of calcein AM to ethidium homodimer demonstrated the degree of cell viability [5,115].…”

Section: Cell Viabilitymentioning

confidence: 99%

“…The fraction of cells which remain adhered or separate from culture substrates after washing reflects the attachment/detachment rate of target cells, which can be generally determined by direct cell counting, spectrophotometric measurement (like MTT and CCK8 assay) and DNA quantification [112,113,116,117,119,120]. In addition, cell morphology and number can be examined by optical microscope, fluorescent staining and SEM imaging, which indirectly explicate the adhesion/deadhesion level of the cells upon a change in temperature [5,35,[115][116][117]121].…”

Section: Cell Attachment and Detachmentmentioning

confidence: 99%

See 2 more Smart Citations

Preparation and Characterization of Thermoresponsive Poly(N-Isopropylacrylamide) for Cell Culture Applications

et al. 2020

View full text Add to dashboard Cite

Poly(N-isopropylacrylamide) (PNIPAAm) is a typical thermoresponsive polymer used widely and studied deeply in smart materials, which is attractive and valuable owing to its reversible and remote "on-off" behavior adjusted by temperature variation. PNIPAAm usually exhibits opposite solubility or wettability across lower critical solution temperature (LCST), and it is readily functionalized making it available in extensive applications. Cell culture is one of the most prospective and representative applications. Active attachment and spontaneous detachment of targeted cells are easily tunable by surface wettability changes and volume phase transitions of PNIPAAm modified substrates with respect to ambient temperature. The thermoresponsive culture platforms and matching thermal-liftoff method can effectively substitute for the traditional cell harvesting ways like enzymatic hydrolysis and mechanical scraping, and will improve the stable and high quality of recovered cells. Therefore, the establishment and detection on PNIPAAm based culture systems are of particular importance. This review covers the important developments and recommendations for future work of the preparation and characterization of temperature-responsive substrates based on PNIPAAm and analogues for cell culture applications.

show abstract

Section: Preparation Of Pnipaam Medicated Substrates For Cell Culturementioning

confidence: 99%

Section: Cell Viabilitymentioning

confidence: 99%

Section: Cell Attachment and Detachmentmentioning

confidence: 99%

See 1 more Smart Citation

Preparation and Characterization of Thermoresponsive Poly(N-Isopropylacrylamide) for Cell Culture Applications

et al. 2020

View full text Add to dashboard Cite

show abstract

“…For the Live/Dead staining, the cells were washed with PBS and treated with 4 × 10 −3 m calcein acetoxymethyl ester (green) and 2 × 10 −3 m ethidium homodimer‐1 (red) at 37 °C for 30 min. [ 32 ] In the case of phalloidin and DAPI staining, the cells were fixed with 4% paraformaldehyde at room temperature for 15 min, permeabilized with 0.2% Triton X‐100 in PBS at 4 °C for 15 min, and blocked with 1% BSA at room temperature for 1 h. Afterward, the cells were stained with TRITC‐conjugated phalloidin and DAPI at room temperature for 60 and 5 min, respectively. For each step, the cells were washed more than three times by PBS for 3 min.…”

Section: Methodsmentioning

confidence: 99%

Metal–Electrolyte Solution Dual‐Mode Electrospinning Process for In Situ Fabrication of Electrospun Bilayer Membrane

Han

Hong

Park

et al. 2020

Adv Materials Inter

Self Cite

View full text Add to dashboard Cite

vivo extracellular matrix (ECM). [2] Various advantages of the electrospun nanofiber membranes, including broad material selection, a high surface-area-to-volume ratio, controllability of physical properties (e.g., diameter and orientation of the nanofibers and porosity and thickness of the membrane), and capability in fabrication of complex nanostructures (e.g., coreshell and Janus nanofibers), [3] have made remarkable advancements in the development of in vitro tissue models and in vivo tissue regeneration. [1,2] Many studies have tried to control the spatial orientation of the electrospun nanofibers through electrospinning, given that the cellular activities, such as adhesion, migration, proliferation, and differentiation, could be promoted by the contact guidance through an anisotropic topographical cue of the electrospun nanofibers. Among various approaches to control the orientation of the electrospun nanofibers, including the electric fieldassisted method, rotating drum method, and magnetic electrospinning, [4-6] the electric field-assisted method readily modulates the orientation of the electrospun nanofibers in a precise and versatile manner. Based on the electric field-assisted method, previous studies have achieved various types of membranes from a uniaxially aligned nanofiber membrane to radially, circumferentially, and biaxially aligned nanofiber membranes, [6-8] showing their potential in manipulating many cell functions, such as myogenic differentiation of myoblast, dural fibroblast migration, and tight junction formation of endothelial cells. [9-11] However, the aligned nanofiber membrane produced by the electric field-assisted method is mechanically unstable, in general, because of its thin (<30 µm thick), anisotropic, and sparsely interconnected nanofibrous structures. [12-14] Hence, the aligned nanofiber membrane is prone to be damaged by external forces occurred with the experimental and surgical manipulations and the aqueous conditions during in vitro and in vivo biomedical applications. [9,15,16] An innovative approach to overcome such limitations of the aligned nanofiber membranes is to develop an electrospun bilayer membrane, also known as a kind of Janus sheet, composed of two membranes, one aligned and the other random, which simultaneously provide the anisotropic topographical Electrospun bilayer membranes comprising two layers, one aligned and the other random, have shown great potential in tissue engineering but previous fabrication processes inevitably relied on manual integration and produced limited types of membranes. Here, a metal-electrolyte solution dual-mode electrospinning (M-ELES) for fabrication of electrospun bilayer membrane based on a metal-electrolyte solution switchable collector is developed. The switchable collector enables random nanofiber deposition directly over the preexisting aligned nanofiber layer in an in situ manner and integration of the layers through an on-demand switch from the metal to the electrolyte solution collector. The electrolyte solution can eff...

show abstract

“…For example, long exposure (more than 20 min) at outside of cell incubator with different temperature, CO 2 concentration, and humidity which can cause negative effect to cell function or viability is required to detach the cells from the PNIPAAm coated surface. 9,10 pH-sensitive property of synthetic material has been also applied for detaching cells without proteolytic cleavage. 11 It has been reported that changing pH condition of human mesenchymal stem cells, that cultured on pH-responsive CaCO 3 precipitated dish, induced cell sheets detachment.…”

Section: Introductionmentioning

confidence: 99%

Alternative method for trypsin-based cell dissociation using poly (amino ester) coating and pH 6.0 PBS

Lee

Kim

et al. 2021

Journal of Bioactive and Compatible Polymers

View full text Add to dashboard Cite

To maintain the cellular functions of a stem cells for therapeutic tissue engineering, an advanced cell culture method for safe cell dissociation is necessary. We developed a novel cell dissociation method by applying pH-responsive bioreducible polymer on the surface of tissue culture plates (TCPs). We applied acid-responsive bioreducible poly (amino ester) (PAE) as a new candidate for surface coating method to develop alternative cell dissociation method against conventional enzyme (trypsin-ethylene diamine tetra acetic acid (EDTA)) treatment. Human adipose derived stem cells (hADSCs) were cultured on and dissociated from PAE-coated TCPs to compare cell adhesion, cell proliferation, cell viability, and functionality to those of the cells cultured on and dissociated with trypsin-EDTA from normal TCPs without PAE coating. To confirm the in vivo therapeutic efficacy of the hADSCs retrieved from PAE-coated TCPs compared to that of the cells retrieved from normal TCPs with trypsin-EDTA, we induced skin defects at the dorsal area of mice and injected the cells collected from both conditions. With the PAE coating method, cell adhesion, cell proliferation, cell viability, and functionality, especially the angiogenic efficacy, were well preserved when compared to those of the cells treated with trypsin-EDTA. In addition to in vitro results, injecting hADSCs retrieved from PAE-coated TCPs showed similar in vivo angiogenesis and wound closing efficiency compared to those of injecting hADSCs retrieved from normal TCPs with trypsin-EDTA treatment at 2 weeks after the transplantation into mouse skin wound models. We proposed the alternative method for the cell dissociation with pH-responsive bioreducible polymer, PAE. This PAE coating method may lead to the development of alternative cell dissociation method without using enzyme for future regenerative medicine and stem cell therapy.

show abstract

Bulk poly(N-isopropylacrylamide) (PNIPAAm) thermoresponsive cell culture platform: toward a new horizon in cell sheet engineering

Abstract: In contrast to the conventional ‘grafting’-based thermoresponsive cell culture platform, we first developed a bulk form of thermoresponsive cell culture platform for attaching/detaching diverse types and origins of the cell sheets in different shape.

Cited by 41 publications

References 61 publications

Preparation and Characterization of Thermoresponsive Poly(N-Isopropylacrylamide) for Cell Culture Applications

Preparation and Characterization of Thermoresponsive Poly(N-Isopropylacrylamide) for Cell Culture Applications

Metal–Electrolyte Solution Dual‐Mode Electrospinning Process for In Situ Fabrication of Electrospun Bilayer Membrane

Alternative method for trypsin-based cell dissociation using poly (amino ester) coating and pH 6.0 PBS

Contact Info

Product

Resources

About