Name: <meta charset="utf8"></head><body>Auteur in de schijnwerpers: een innovatieve celschraper ontwerpen met behulp van interactie-ervaringsontwerp en analyse van gebruikersvereisten
Uploaded: 2024-10-04T12:30:00
Description: The reliability of cellular migration measurement in wound healing assays is frequently undermined by the prevalent methodological instability, i.e., ...

<meta charset="utf8"></head><body>Innoveren van een verbeterde celschraper voor celbeschadiging door analyse van gebruikersvereisten

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>CCK-8 Kit</td>
			<td>Beyotime Company, China</td>
			<td>C0037</td>
			<td></td>
		</tr>
		<tr>
			<td>digital microscope system</td>
			<td>Olympus</td>
			<td>IX81</td>
			<td></td>
		</tr>
		<tr>
			<td>fetal bovine serum</td>
			<td>Gibco, USA</td>
			<td>16000044</td>
			<td></td>
		</tr>
		<tr>
			<td>HOS&#160;</td>
			<td>Procell Life Science &#38; Technology Co., Ltd</td>
			<td>CL-0360</td>
			<td></td>
		</tr>
		<tr>
			<td>Image-Pro Plus</td>
			<td>Media Cybernetics</td>
			<td>version 6.0</td>
			<td></td>
		</tr>
		<tr>
			<td>KeyShot</td>
			<td>Luxion</td>
			<td>version 11.0</td>
			<td>3D rendering software</td>
		</tr>
		<tr>
			<td>microplate reader</td>
			<td>BioTek, German</td>
			<td>ELX808</td>
			<td></td>
		</tr>
		<tr>
			<td>Minimum Essential Medium</td>
			<td>Gibco, USA</td>
			<td>11095080</td>
			<td></td>
		</tr>
		<tr>
			<td>Pantone matching system</td>
			<td>Pantone</td>
			<td></td>
			<td>commercial color matching</td>
		</tr>
		<tr>
			<td>penicillin-streptomycin</td>
			<td>Beyotime Company, China</td>
			<td>ST488</td>
			<td></td>
		</tr>
		<tr>
			<td>Photoshop</td>
			<td>Adobe</td>
			<td></td>
			<td>photo and design software</td>
		</tr>
		<tr>
			<td>Rhinoceros 3D</td>
			<td>Robert McNeel &#38; Associates</td>
			<td>version 7.0</td>
			<td>3D design software</td>
		</tr>
		<tr>
			<td>TC20 Automated Cell Counter</td>
			<td>Bio-Rad</td>
			<td>TC20</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin</td>
			<td>Cytiva&#160;HyClone, United State</td>
			<td>SH30042.01</td>
			<td></td>
		</tr>
	</tbody>
</table>

a tool to automatically create stable and reproducible cell-free gaps for improving the reliability of cell wound healing assay

Tumors are characterized by distinct hallmarks such as selective growth advantages, metabolic rewiring, and immune modulation, all of which intriguingly contribute to enhanced cell migration, a critical malignant behavior of tumor cells. This feature directly affects the distant metastasis of the primary tumor, compromising the long-term survival of patients1,2,3. Selective growth advantages enable cancer cells to outcompete normal cells, while metabolic rewiring supports this rapid proliferation by altering energy pathways. Concurrently, immune modulation allows tumors to evade the body&#39;s defenses. Studies underscore the severity of this issue, showing that lung metastasis, often a result of enhanced cell migration, is a terminal event leading to the death of patients with various cancers4. For instance, breast cancers5, cervical carcinoma4, and osteosarcoma6 account for 20%, 9%, and 30% of such cases, respectively. Therefore, assessing tumor cell migration has become integral to current oncology research, further highlighting the multifaceted nature of tumor progression.
Cell wound healing assay is an easy-to-use method for measuring in vitro cellular migration, often employed in oncological studies7. Most experimenters use pipette tips to create cell wounds manually8. Although such a method could form cell wounds rapidly and conveniently, it still has many limitations that affect reproducibility and accuracy for evaluating cell migration. Firstly, using pipette tips to create scratches manually is highly influenced by the operator&#39;s operating angle, force, and speed, affecting the method&#39;s repeatability8. Secondly, tip-generated cell defects usually have jagged edges rather than straight edges because pipette tips are plastic products with certain elasticity9. Some studies generate wounds by placing prefabricated culture inserts directly into the cell culture plate to restrict the range of cell proliferation10. This approach circumvents the limitation of the tips-based method, such as jagged edges and reproducibility. However, even with biocompatible materials, the long-term coexistence of the embedding with the cells still impacts cell growth11. Moreover, the embedding may also cause cellular epigenetic changes in the marginal zone due to contact restriction12. Also, contact-insert produced from biocompatible materials are expensive and difficult to reuse, limiting their feasibility13. Therefore, a novel, reproducible, and practical tool is needed to easily quantify in vitro cellular migration.
The primary goal of this method is to introduce an innovative tool for quantifying in vitro cellular migration in oncological studies, addressing the limitations of existing techniques and enhancing reproducibility and accuracy in assessing cellular migration.
The rationale behind this technique&#39;s development lies in the critical importance of evaluating tumor cell migration in oncology. Tumors exhibit distinctive hallmarks, including selective growth advantages, metabolic rewiring, and immune modulation, all contributing to enhanced cell migration, a fundamental aspect of cancer malignancy. This method aims to provide a more dependable means of studying cellular migration, contributing to a deeper understanding of tumor behavior.
This method offers substantial advantages over existing techniques. Manual scratch assays can suffer from operator-dependent interference, while culture inserts may impact cell growth and gene expression. In contrast, this method offers improved repeatability, accuracy, and practicality, presenting a cost-effective solution for measuring in vitro cellular migration in oncology research. It addresses a crucial need for a reliable and accessible tool to study cell migration in various cancer types, making it a valuable addition to the field.

<meta charset="utf8"></head><body>Auteur in de schijnwerpers: een innovatieve celschraper ontwerpen met behulp van interactie-ervaringsontwerp en analyse van gebruikersvereisten

Een hulpmiddel om automatisch stabiele en reproduceerbare celvrije openingen te creëren voor het verbeteren van de betrouwbaarheid van de celwondgenezingstest

Through the incorporation of interaction experience design and user requirements analysis, we introduce a innovative cell scraper that enhance cellular wound healing assays in terms of reproducibility, dependability, practicality, cellular integrity, and user experiences. We authorized consultative rhinal, keystone, and photocell for product visualizations. This approach that only is in pre-producing the actual product, but also we enhance communications among team members.Additionally, it contributes to reducing the overall year costs. In experimental validations, the cell scraper showed higher survival and proliferation risk than the TIPS group post wound completion confirmed by an EdU assay. This indicates a scraper superiority in creating stable biologically reproducible cell walls compared to the traditional method with minimal impact on cell health.

The reliability of cellular migration measurement in wound healing assays is frequently undermined by the prevalent methodological instability, i.e., ...

a-tool-to-automatically-create-stable-reproducible-cell-free-gaps-for

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Research

A Tool to Automatically Create Stable and Reproducible Cell-free Gaps for Improving the Reliability of Cell Wound Healing Assay

JoVE Journal

Biology

The reliability of cellular migration measurement in wound healing assays is frequently undermined by the prevalent methodological instability, i.e., tip-based method. We introduce an innovative instrument designed to address these limitations. Our novel cell scraper surpasses the current approach, generating a more consistent and stable cell-free gap. Repeated biological experiments reveal that the cell-free gap produced by the cell scraper exhibits straighter edges and uniform size and shape compared to the tip-based technique (p &lt; 0.05). In terms of product design, the cell scraper boasts a refined color scheme suited for laboratory environments, enhancing the monitoring of experimental outcomes, and permits sterilization through autoclaving for reuse. Notably, after treatment, the cell scraper demonstrates a negligible effect on cell viability and proliferation (97.31% and 24.41%, respectively). Conversely, the tip-based method yields lower cell viability (91.37%) and proliferation (18.79%). This investigation presents the cell scraper as a novel, reusable device capable of generating reproducible cell-free gaps while preserving cellular viability, thereby augmenting the reliability of wound healing assays in comparison to existing techniques.

Through the incorporation of interaction experience design and user requirements analysis, we introduce an innovative cell scraper that enhances cellular wound healing assays in terms of reproducibility, dependability, practicality, cellular integrity, and user experience.

Questionnaire Survey and User Requirement Analysis for Designing Innovative Cell Wounding Tools

To investigate the requirements of the user community, deliver questionnaires to biologists or experimentalists working on cell wound healing assays. Examples of a few relevant questions are displayed on the screen. After collecting the completed questionnaires, proceed to use them for the study.Investigate the respondent's perceptions of tools such as pipette tips, and cell culture inserts in cell wound healing experiments. Using the means and chain theory based on laddering technique in the questionnaire section, understand and gather their reasons for selecting one tool over another. The employment of the hard laddering method to analyze the requirements of users from 97 fully completed questionnaires, showed that the most troubling problems in cell wound healing assay were serrated edge and unstable width, or areas of wounds.Other notable results showed that for creating wounds, the use of pipette tips was much more prevalent than cell culture inserts. It was mainly ascribed to the high cost of cell culture inserts. As for the innovative tool to generate cell wounds, 86.6%of respondents were optimistic.

Vragenlijstonderzoek en analyse van de gebruikersbehoefte voor het ontwerpen van innovatieve hulpmiddelen voor celwonden

Designing and 3D Modeling of Innovative Cell Wounding Tools Based on User Requirement Analysis and Assessing Their Cell Wounding Potential

To begin, draw a sketch from the insights gathered from the questionnaire survey to initiate the design process. Use this preliminary sketch as the foundational blueprint. Using the modeling software, elaborate on the dimensions and layout of each component by applying the DEEM, DEEM radius and DEEM diameter functions.Next, utilizing a vernier caliper with a precision of 0.02 millimeters measure actual six well plates to ascertain and confirm that the final dimensions of the cell scraper match the values mentioned on the screen. Pay attention to the details, especially the product height and fitness in the well to facilitate a smoother assembly. Begin the 3D model designing in the software by creating a basic model.Click buttons like extrude CRV for stretching, and loft for shaping the design. After refining the model, then click filet edge for smooth edges. Import the model into a 3D rendering software.Apply materials including plastic, sponge, and steel. Then adjust the lighting for realistic rendering. Finally, in photo and design software, enhance the images, contrast and saturation, and add background elements for context to ensure a high quality realistic product representation.After the scraper is manufactured, autoclave it at 121.3 degrees Celsius for 30 minutes to ensure it retains its original shape and physical properties. For the tip based wounding method, use a 100 microliter tip and drag it across the medium with one stroke in the horizontal direction, and another in the vertical direction. For the cell scraper based wounding method, place the scraper prototype in one of the wells and press once gently with the help of the tweezer.A three-dimensional model of the cell scraper was constructed using the 3D design software. The linkage lever inside the cell scraper was connected to the top button and the end move downward after applying pressure to make the end buckle. The spring inside the slide was released to push the tips to move.Finally, it stopped at the sponge with a narrow slit to finish the scraping and then toggle to reset when used again. The cell scraper integrated excellently with six well plates, the upper cross shaped holder fitted closely to the edge of the plate to provide a strong fixation. The tips also fitted precisely to the bottom, facilitating the smooth generation of cell-free areas.The cell scraper held its original shape and performance, even after autoclaving. The cell scraper outperformed the existing cell wound creation method in the benchmark test. After repeated tests, the cell scratcher showed high stability and reproducibility in the preparation of the wound, and presented a low fluctuation in the area of each wound and nearly straight edges.In contrast, the area of each wound prepared by the pipette tips showed a higher fluctuation and deviation to straight edges.

Ontwerpen en 3D-modelleren van innovatieve hulpmiddelen voor celwonden op basis van analyse van gebruikersvereisten en beoordeling van hun potentieel voor celwonden