Efficient Generation of Chondrocytes From Bone Marrow–Derived Mesenchymal Stem Cells in a 3D Culture System: Protocol for a Practical Model for Assessing Anti-Inflammatory Therapies
BackgroundChondrocytes are the primary cells responsible for maintaining cartilage integrity and function. Their role in cartilage homeostasis and response to inflammation is crucial for understanding the progression and potential therapeutic interventions for various cartila...
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| Format: | Article |
| Language: | English |
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JMIR Publications
2023-07-01
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| Series: | JMIR Research Protocols |
| Online Access: | https://www.researchprotocols.org/2023/1/e42964 |
| _version_ | 1797733961739272192 |
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| author | Rajashree Patnaik Shirin Jannati Bala Mohan Sivani Manfredi Rizzo Nerissa Naidoo Yajnavalka Banerjee |
| author_facet | Rajashree Patnaik Shirin Jannati Bala Mohan Sivani Manfredi Rizzo Nerissa Naidoo Yajnavalka Banerjee |
| author_sort | Rajashree Patnaik |
| collection | DOAJ |
| description |
BackgroundChondrocytes are the primary cells responsible for maintaining cartilage integrity and function. Their role in cartilage homeostasis and response to inflammation is crucial for understanding the progression and potential therapeutic interventions for various cartilage-related disorders. Developing an accessible and cost-effective model to generate viable chondrocytes and to assess their response to different bioactive compounds can significantly advance our knowledge of cartilage biology and contribute to the discovery of novel therapeutic approaches.
ObjectiveWe developed a novel, streamlined protocol for generating chondrocytes from bone marrow–derived mesenchymal stem cells (BMSCs) in a 3D culture system that offers significant implications for the study of cartilage biology and the discovery of potential therapeutic interventions for cartilage-related and associated disorders.
MethodsWe developed a streamlined protocol for generating chondrocytes from BMSCs in a 3D culture system using an “in-tube” culture approach. This simple pellet-based 3D culture system allows for cell aggregation and spheroid formation, facilitating cell-cell and cell–extracellular matrix interactions that better mimic the in vivo cellular environment compared with 2D monolayer cultures. A proinflammatory chondrocyte model was created by treating the chondrocytes with lipopolysaccharide and was subsequently used to evaluate the anti-inflammatory effects of vitamin D, curcumin, and resveratrol.
ResultsThe established protocol successfully generated a large quantity of viable chondrocytes, characterized by alcian blue and toluidine blue staining, and demonstrated versatility in assessing the anti-inflammatory effects of various bioactive compounds. The chondrocytes exhibited reduced inflammation, as evidenced by the decreased tumor necrosis factor-α levels, in response to vitamin D, curcumin, and resveratrol treatment.
ConclusionsOur novel protocol offers an accessible and cost-effective approach for generating chondrocytes from BMSCs and for evaluating potential therapeutic leads in the context of inflammatory chondrocyte–related diseases. Although our approach has several advantages, further investigation is required to address its limitations, such as the potential differences between chondrocytes generated using our protocol and those derived from other established methods, and to refine the model for broader applicability and clinical translation. |
| first_indexed | 2024-03-12T12:36:11Z |
| format | Article |
| id | doaj.art-5889e7fd36a1489f81087b14c09a57b3 |
| institution | Directory Open Access Journal |
| issn | 1929-0748 |
| language | English |
| last_indexed | 2024-03-12T12:36:11Z |
| publishDate | 2023-07-01 |
| publisher | JMIR Publications |
| record_format | Article |
| series | JMIR Research Protocols |
| spelling | doaj.art-5889e7fd36a1489f81087b14c09a57b32023-08-29T00:08:27ZengJMIR PublicationsJMIR Research Protocols1929-07482023-07-0112e4296410.2196/42964Efficient Generation of Chondrocytes From Bone Marrow–Derived Mesenchymal Stem Cells in a 3D Culture System: Protocol for a Practical Model for Assessing Anti-Inflammatory TherapiesRajashree Patnaikhttps://orcid.org/0000-0003-2654-7007Shirin Jannatihttps://orcid.org/0000-0002-8058-3204Bala Mohan Sivanihttps://orcid.org/0000-0002-8631-3926Manfredi Rizzohttps://orcid.org/0000-0002-9549-8504Nerissa Naidoohttps://orcid.org/0000-0002-0924-8796Yajnavalka Banerjeehttps://orcid.org/0000-0002-7546-8893 BackgroundChondrocytes are the primary cells responsible for maintaining cartilage integrity and function. Their role in cartilage homeostasis and response to inflammation is crucial for understanding the progression and potential therapeutic interventions for various cartilage-related disorders. Developing an accessible and cost-effective model to generate viable chondrocytes and to assess their response to different bioactive compounds can significantly advance our knowledge of cartilage biology and contribute to the discovery of novel therapeutic approaches. ObjectiveWe developed a novel, streamlined protocol for generating chondrocytes from bone marrow–derived mesenchymal stem cells (BMSCs) in a 3D culture system that offers significant implications for the study of cartilage biology and the discovery of potential therapeutic interventions for cartilage-related and associated disorders. MethodsWe developed a streamlined protocol for generating chondrocytes from BMSCs in a 3D culture system using an “in-tube” culture approach. This simple pellet-based 3D culture system allows for cell aggregation and spheroid formation, facilitating cell-cell and cell–extracellular matrix interactions that better mimic the in vivo cellular environment compared with 2D monolayer cultures. A proinflammatory chondrocyte model was created by treating the chondrocytes with lipopolysaccharide and was subsequently used to evaluate the anti-inflammatory effects of vitamin D, curcumin, and resveratrol. ResultsThe established protocol successfully generated a large quantity of viable chondrocytes, characterized by alcian blue and toluidine blue staining, and demonstrated versatility in assessing the anti-inflammatory effects of various bioactive compounds. The chondrocytes exhibited reduced inflammation, as evidenced by the decreased tumor necrosis factor-α levels, in response to vitamin D, curcumin, and resveratrol treatment. ConclusionsOur novel protocol offers an accessible and cost-effective approach for generating chondrocytes from BMSCs and for evaluating potential therapeutic leads in the context of inflammatory chondrocyte–related diseases. Although our approach has several advantages, further investigation is required to address its limitations, such as the potential differences between chondrocytes generated using our protocol and those derived from other established methods, and to refine the model for broader applicability and clinical translation.https://www.researchprotocols.org/2023/1/e42964 |
| spellingShingle | Rajashree Patnaik Shirin Jannati Bala Mohan Sivani Manfredi Rizzo Nerissa Naidoo Yajnavalka Banerjee Efficient Generation of Chondrocytes From Bone Marrow–Derived Mesenchymal Stem Cells in a 3D Culture System: Protocol for a Practical Model for Assessing Anti-Inflammatory Therapies JMIR Research Protocols |
| title | Efficient Generation of Chondrocytes From Bone Marrow–Derived Mesenchymal Stem Cells in a 3D Culture System: Protocol for a Practical Model for Assessing Anti-Inflammatory Therapies |
| title_full | Efficient Generation of Chondrocytes From Bone Marrow–Derived Mesenchymal Stem Cells in a 3D Culture System: Protocol for a Practical Model for Assessing Anti-Inflammatory Therapies |
| title_fullStr | Efficient Generation of Chondrocytes From Bone Marrow–Derived Mesenchymal Stem Cells in a 3D Culture System: Protocol for a Practical Model for Assessing Anti-Inflammatory Therapies |
| title_full_unstemmed | Efficient Generation of Chondrocytes From Bone Marrow–Derived Mesenchymal Stem Cells in a 3D Culture System: Protocol for a Practical Model for Assessing Anti-Inflammatory Therapies |
| title_short | Efficient Generation of Chondrocytes From Bone Marrow–Derived Mesenchymal Stem Cells in a 3D Culture System: Protocol for a Practical Model for Assessing Anti-Inflammatory Therapies |
| title_sort | efficient generation of chondrocytes from bone marrow derived mesenchymal stem cells in a 3d culture system protocol for a practical model for assessing anti inflammatory therapies |
| url | https://www.researchprotocols.org/2023/1/e42964 |
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