Development of Low-Cost CNC-Milled PMMA Microfluidic Chips as a Prototype for Organ-on-a-Chip and Neurospheroid Applications

Improved in vitro models are needed to reduce costs and delays in central nervous system (CNS) drug discovery. The FDA Modernization Acts 2.0 and 3.0 require human-centered alternative testing methods to mitigate animal-based experiments and discovery delays, and to ensure human safety. Developing c...

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Main Authors: Sushmita Mishra, Ginia Mondal, Murali Kumarasamy
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Organoids
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Online Access:https://www.mdpi.com/2674-1172/4/2/13
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author Sushmita Mishra
Ginia Mondal
Murali Kumarasamy
author_facet Sushmita Mishra
Ginia Mondal
Murali Kumarasamy
author_sort Sushmita Mishra
collection DOAJ
description Improved in vitro models are needed to reduce costs and delays in central nervous system (CNS) drug discovery. The FDA Modernization Acts 2.0 and 3.0 require human-centered alternative testing methods to mitigate animal-based experiments and discovery delays, and to ensure human safety. Developing cost-efficient, flexible microfluidic chips is essential to advance organ-on-chip (OoC) technology for drug discovery and disease modeling. While CNC micromilling shows promise for fabricating microfluidic devices, it remains underutilized due to limited accessibility. We present a simple CNC-milled flexible microfluidic chip fabricated from thermoplastic poly (methyl methacrylate) (PMMA). The structure of the microplate included drilled openings for connecting the wells. The chip’s biocompatibility was evaluated with isolated primary neuronal cultures from postnatal Wistar rat pups (p1). Primary cells cultured in the device showed high viability, differentiation, and 3D neurosphere formation, similar to conventional well-plate cultures. Neuronal cultures showed neurite growth and functional markers. Although cleanroom-based methods provide higher accuracy, the chip effectively promotes cell viability, differentiation, and alignment, offering an ideal platform for tissue modeling and OoC applications. It allows cell biologists to quickly create prototypes at lower cost and in less time than required for soft lithography and is a viable alternative to the current manufacturing methods.
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spelling doaj-art-46bef30aaa914e13a8ba39ab41b013642025-08-20T03:16:21ZengMDPI AGOrganoids2674-11722025-06-01421310.3390/organoids4020013Development of Low-Cost CNC-Milled PMMA Microfluidic Chips as a Prototype for Organ-on-a-Chip and Neurospheroid ApplicationsSushmita Mishra0Ginia Mondal1Murali Kumarasamy2Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Hajipur, Export Promotion Industrial Park (EPIP), Vaishali 844102, Bihar, IndiaDepartment of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Hajipur, Export Promotion Industrial Park (EPIP), Vaishali 844102, Bihar, IndiaDepartment of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Hajipur, Export Promotion Industrial Park (EPIP), Vaishali 844102, Bihar, IndiaImproved in vitro models are needed to reduce costs and delays in central nervous system (CNS) drug discovery. The FDA Modernization Acts 2.0 and 3.0 require human-centered alternative testing methods to mitigate animal-based experiments and discovery delays, and to ensure human safety. Developing cost-efficient, flexible microfluidic chips is essential to advance organ-on-chip (OoC) technology for drug discovery and disease modeling. While CNC micromilling shows promise for fabricating microfluidic devices, it remains underutilized due to limited accessibility. We present a simple CNC-milled flexible microfluidic chip fabricated from thermoplastic poly (methyl methacrylate) (PMMA). The structure of the microplate included drilled openings for connecting the wells. The chip’s biocompatibility was evaluated with isolated primary neuronal cultures from postnatal Wistar rat pups (p1). Primary cells cultured in the device showed high viability, differentiation, and 3D neurosphere formation, similar to conventional well-plate cultures. Neuronal cultures showed neurite growth and functional markers. Although cleanroom-based methods provide higher accuracy, the chip effectively promotes cell viability, differentiation, and alignment, offering an ideal platform for tissue modeling and OoC applications. It allows cell biologists to quickly create prototypes at lower cost and in less time than required for soft lithography and is a viable alternative to the current manufacturing methods.https://www.mdpi.com/2674-1172/4/2/13computer numerical control (CNC) machineorgan-on-a-chipmicromillingprimary neural cells3D neurospheroid
spellingShingle Sushmita Mishra
Ginia Mondal
Murali Kumarasamy
Development of Low-Cost CNC-Milled PMMA Microfluidic Chips as a Prototype for Organ-on-a-Chip and Neurospheroid Applications
Organoids
computer numerical control (CNC) machine
organ-on-a-chip
micromilling
primary neural cells
3D neurospheroid
title Development of Low-Cost CNC-Milled PMMA Microfluidic Chips as a Prototype for Organ-on-a-Chip and Neurospheroid Applications
title_full Development of Low-Cost CNC-Milled PMMA Microfluidic Chips as a Prototype for Organ-on-a-Chip and Neurospheroid Applications
title_fullStr Development of Low-Cost CNC-Milled PMMA Microfluidic Chips as a Prototype for Organ-on-a-Chip and Neurospheroid Applications
title_full_unstemmed Development of Low-Cost CNC-Milled PMMA Microfluidic Chips as a Prototype for Organ-on-a-Chip and Neurospheroid Applications
title_short Development of Low-Cost CNC-Milled PMMA Microfluidic Chips as a Prototype for Organ-on-a-Chip and Neurospheroid Applications
title_sort development of low cost cnc milled pmma microfluidic chips as a prototype for organ on a chip and neurospheroid applications
topic computer numerical control (CNC) machine
organ-on-a-chip
micromilling
primary neural cells
3D neurospheroid
url https://www.mdpi.com/2674-1172/4/2/13
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AT muralikumarasamy developmentoflowcostcncmilledpmmamicrofluidicchipsasaprototypefororganonachipandneurospheroidapplications