Simple and Cost-Effective Generation of 3D Cell Sheets and Spheroids Using Curvature-Controlled Paraffin Wax Substrates

Abstract The challenges associated with animal testing in pharmaceutical development have driven the search for alternative in vitro models that mimic human tissues more accurately. In this study, we present a simple and cost-effective method for generating 3D cell sheets and spheroids using curvatu...

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Main Authors: Huijung Kim, Kyeong-Mo Koo, Chang-Dae Kim, Min Ji Byun, Chun Gwon Park, Hyungbin Son, Hyung-Ryong Kim, Tae-Hyung Kim
Format: Article
Language:English
Published: SpringerOpen 2024-10-01
Series:Nano Convergence
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Online Access:https://doi.org/10.1186/s40580-024-00451-4
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author Huijung Kim
Kyeong-Mo Koo
Chang-Dae Kim
Min Ji Byun
Chun Gwon Park
Hyungbin Son
Hyung-Ryong Kim
Tae-Hyung Kim
author_facet Huijung Kim
Kyeong-Mo Koo
Chang-Dae Kim
Min Ji Byun
Chun Gwon Park
Hyungbin Son
Hyung-Ryong Kim
Tae-Hyung Kim
author_sort Huijung Kim
collection DOAJ
description Abstract The challenges associated with animal testing in pharmaceutical development have driven the search for alternative in vitro models that mimic human tissues more accurately. In this study, we present a simple and cost-effective method for generating 3D cell sheets and spheroids using curvature-controlled paraffin wax films, which are easily accessible laboratory materials that eliminate the need for extracellular matrix (ECM) components or thermo-responsive polymers. By adjusting the curvature of the paraffin wax film, we successfully generated human periodontal ligament fibroblast (HPdLF) cell sheets and bone marrow-derived mesenchymal stem cell (hBMSC) spheroids. Key parameters, such as cell density, substrate curvature, and incubation time, were identified as critical factors for optimizing the formation of these 3D structures. In addition, the use of quantum dots (QDs) for cell tracking enabled long-term visualization and distinction between different cell types within complex tissue-like structures. We further demonstrated that wrapping the hBMSC spheroids with HPdLF cell sheets partially replicated the connective tissue structure of the periodontal ligament surrounding the tooth root. This highlights the potential of this platform for the construction of more sophisticated tissue-mimicking assemblies. In conclusion, curvature-controlled paraffin wax films provide a versatile and practical approach for 3D cell cultures. This simplifies the generation of both cell sheets and spheroids, offering a promising tool for tissue engineering and regenerative medicine applications, where precise cell-to-cell interactions are essential. Graphical abstract
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spelling doaj-art-e9141bb3a6ce462b9d2464ca28c908752025-08-20T02:18:25ZengSpringerOpenNano Convergence2196-54042024-10-0111111210.1186/s40580-024-00451-4Simple and Cost-Effective Generation of 3D Cell Sheets and Spheroids Using Curvature-Controlled Paraffin Wax SubstratesHuijung Kim0Kyeong-Mo Koo1Chang-Dae Kim2Min Ji Byun3Chun Gwon Park4Hyungbin Son5Hyung-Ryong Kim6Tae-Hyung Kim7Department of Biomedical Engineering, Institute for Cross-Disciplinary Studies (ICS), Sungkyunkwan University (SKKU)Department of Biomedical Engineering, Institute for Cross-Disciplinary Studies (ICS), Sungkyunkwan University (SKKU)Department of Biomedical Engineering, Institute for Cross-Disciplinary Studies (ICS), Sungkyunkwan University (SKKU)Department of Biomedical Engineering, Institute for Cross-Disciplinary Studies (ICS), Sungkyunkwan University (SKKU)Department of Biomedical Engineering, Institute for Cross-Disciplinary Studies (ICS), Sungkyunkwan University (SKKU)School of Integrative Engineering, Chung-Ang UniversityDepartment of Pharmacology, College of Dentistry, Jeonbuk National UniversityDepartment of Biomedical Engineering, Institute for Cross-Disciplinary Studies (ICS), Sungkyunkwan University (SKKU)Abstract The challenges associated with animal testing in pharmaceutical development have driven the search for alternative in vitro models that mimic human tissues more accurately. In this study, we present a simple and cost-effective method for generating 3D cell sheets and spheroids using curvature-controlled paraffin wax films, which are easily accessible laboratory materials that eliminate the need for extracellular matrix (ECM) components or thermo-responsive polymers. By adjusting the curvature of the paraffin wax film, we successfully generated human periodontal ligament fibroblast (HPdLF) cell sheets and bone marrow-derived mesenchymal stem cell (hBMSC) spheroids. Key parameters, such as cell density, substrate curvature, and incubation time, were identified as critical factors for optimizing the formation of these 3D structures. In addition, the use of quantum dots (QDs) for cell tracking enabled long-term visualization and distinction between different cell types within complex tissue-like structures. We further demonstrated that wrapping the hBMSC spheroids with HPdLF cell sheets partially replicated the connective tissue structure of the periodontal ligament surrounding the tooth root. This highlights the potential of this platform for the construction of more sophisticated tissue-mimicking assemblies. In conclusion, curvature-controlled paraffin wax films provide a versatile and practical approach for 3D cell cultures. This simplifies the generation of both cell sheets and spheroids, offering a promising tool for tissue engineering and regenerative medicine applications, where precise cell-to-cell interactions are essential. Graphical abstracthttps://doi.org/10.1186/s40580-024-00451-4Paraffin waxCell sheetSpheroidsPeriodontal ligament cellMesenchymal stem cellQuantum dots
spellingShingle Huijung Kim
Kyeong-Mo Koo
Chang-Dae Kim
Min Ji Byun
Chun Gwon Park
Hyungbin Son
Hyung-Ryong Kim
Tae-Hyung Kim
Simple and Cost-Effective Generation of 3D Cell Sheets and Spheroids Using Curvature-Controlled Paraffin Wax Substrates
Nano Convergence
Paraffin wax
Cell sheet
Spheroids
Periodontal ligament cell
Mesenchymal stem cell
Quantum dots
title Simple and Cost-Effective Generation of 3D Cell Sheets and Spheroids Using Curvature-Controlled Paraffin Wax Substrates
title_full Simple and Cost-Effective Generation of 3D Cell Sheets and Spheroids Using Curvature-Controlled Paraffin Wax Substrates
title_fullStr Simple and Cost-Effective Generation of 3D Cell Sheets and Spheroids Using Curvature-Controlled Paraffin Wax Substrates
title_full_unstemmed Simple and Cost-Effective Generation of 3D Cell Sheets and Spheroids Using Curvature-Controlled Paraffin Wax Substrates
title_short Simple and Cost-Effective Generation of 3D Cell Sheets and Spheroids Using Curvature-Controlled Paraffin Wax Substrates
title_sort simple and cost effective generation of 3d cell sheets and spheroids using curvature controlled paraffin wax substrates
topic Paraffin wax
Cell sheet
Spheroids
Periodontal ligament cell
Mesenchymal stem cell
Quantum dots
url https://doi.org/10.1186/s40580-024-00451-4
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