Topographical evaluation of human mesenchymal stem cells during osteogenic differentiation using scanning ion conductance microscopy

Nozawa, Kota; Zhang, Xuyang; Nakamura, Tokiko; Nashimoto, Yuji; Takahashi, Yasufumi; Ino, Kosuke; Shiku, Hitoshi

doi:10.1016/j.electacta.2023.142192

Cited by 4 publications

(1 citation statement)

References 30 publications

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“…An understanding of the response of the same A549 cells to cisplatin over time is necessary for studying dynamic morphological changes. SICM allows for continuous topography measurement over several days, and has been reported for the studies of stem cells during osteogenic differentiation, 69 where individual images can be recorded relatively quickly, in the course of a few minutes. 70 We first performed a 24 h control measurement to confirm that the SICM imaging environment does not alter the native morphology of a single cancer cell over time, where we observed no changes in morphology (Fig.…”

Section: Resultsmentioning

confidence: 99%

Scanning ion conductance microscopy revealed cisplatin-induced morphological changes related to apoptosis in single adenocarcinoma cells

Muhammed,

Lazenby

2024

Anal. Methods

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Section: Resultsmentioning

confidence: 99%

Scanning ion conductance microscopy revealed cisplatin-induced morphological changes related to apoptosis in single adenocarcinoma cells

Muhammed,

Lazenby

2024

Anal. Methods

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Scanning gel electrochemical microscopy (SGECM): Elaboration and cross-linking of chitosan-based gel probes

Dang,

Echeveste Salazar,

Walcarius

et al. 2024

Electrochimica Acta

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Label-Free Imaging of Mesenchymal Stem Cell Spheroid Differentiation with Flexible-Probe SECM and a Microfluidic Device

Zhao,

Wang,

Huang

et al. 2024

Anal. Chem.

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Mesenchymal stem cells (MSCs) have emerged as an indispensable source for stem cell research and preclinical studies due to their capacity for in vitro proliferation and their potential to differentiate into mesodermal lineages, particularly into osteoblasts. This capability has propelled their application in the fields of bone regeneration and osteochondral repair. Traditional methodologies for assessing the differentiation status of MSCs necessitate invasive procedures such as cell lysis or fixation. In this study, we introduce a nondestructive technique that utilizes an integrated label-free approach to evaluate the osteogenic maturation of MSC spheroid aggregates. This method employs scanning electrochemical microscopy (SECM) with a flexible probe in conjunction with a top-removable microfluidic device designed for easy SECM access. By tracking the production rate of p-aminophenol (PAP) in the generation/collection mode and assessing morphological changes via the negative feedback mode using [Ru(NH3)6]Cl3 (Ruhex), we can discern variations in the alkaline phosphatase (ALP) activity indicative of osteogenic differentiation. This innovative strategy enables the direct evaluation of osteogenic differentiation in MSC spheroids cultured within microwell arrays without necessitating any labeling procedures. The utilization of a flexible microelectrode as the probe that scans in contact mode (with probe–substrate distances potentially as minimal as 0 μm) affords enhanced resolution compared to the traditional stiff-probe technique. Furthermore, this method is compatible with subsequent molecular biology assays, including gene expression analysis and immunofluorescence, thereby confirming the electrochemical findings and establishing the validity of this integrative approach.

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Topographical evaluation of human mesenchymal stem cells during osteogenic differentiation using scanning ion conductance microscopy

Cited by 4 publications

References 30 publications

Scanning ion conductance microscopy revealed cisplatin-induced morphological changes related to apoptosis in single adenocarcinoma cells

Scanning ion conductance microscopy revealed cisplatin-induced morphological changes related to apoptosis in single adenocarcinoma cells

Scanning gel electrochemical microscopy (SGECM): Elaboration and cross-linking of chitosan-based gel probes

Label-Free Imaging of Mesenchymal Stem Cell Spheroid Differentiation with Flexible-Probe SECM and a Microfluidic Device

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