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Cellular and Molecular Bioengineering

Cellular and Molecular Bioengineering - Poster Session A

Poster H15 - A microfluidic device to quantify immune cell transmigration in the vasculature

Thursday, October 24, 2024

10:00 AM – 11:00 AM EST

Location: Exhibit Hall E, F & G

Presenting Author(s)

GI

Gulimina Imran

Undergraduate Capstone Researcher
New Jersey Institute of Technology, United States

Co-Author(s)

SO

Sahmet Ozda

Undergraduate Capstone Researcher
New Jersey Institute of Technology, United States
Theresa Carlos (she/her/hers)

Undergraduate Capstone Researcher
New Jersey Institute of Technology
Newark, New Jersey, United States
AV

Ana Villanueva

Undergraduate Capstone Researcher
New Jersey Institute of Technology, United States
CG

Clara Ghaly

Undergraduate Capstone Researcher
New Jersey Institute of Technology, United States
SG

Subham Guin, MS (he/him/his)

Ph.D. Student
New Jersey Institute of Technology
Newark, New Jersey, United States

Presenting Author(s)

Alexander Buffone, Jr., PhD (he/him/his)

Assistant Professor
New Jersey Institute of Technology
Newark, New Jersey, United States

Introduction: In this project, we will be fabricating a passive microfluidic device that utilizes channel geometry and size to manipulate the flow of cells through a transmembrane by utilizing a technique known as transwell assay. The microfluidic device will consist of 2 photopolymer resin plates with their own separate well vessel/channel with unidirectional flow channels and a membrane in the middle separating the two. The top plate will consist of a cultured fluid insert of neutrophils subjected to shear flow, while the bottom plate will consist of a fluid with chemoattractants subjected to a shear flow as well. By simulating physiological conditions, this device aims to provide insights into cell migration dynamics, particularly how cells interact with and traverse across porous barriers.Our microfluidic device will be used to study cell migration in response to potential therapeutic agents or conditions, thus facilitating the development of targeted interventions for various diseases.

Materials and

Methods: The materials to be utilized in fabricating the microfluidic device will include clear photopolymer resin for 3D printing the plates and channels. Additionally, a circular membrane with a specific coating to facilitate cell attachment will be employed. Screws and gasket material will be used for assembly to prevent potential leakage, while tubing sets will be utilized to establish connections between the microfluidic device channels and the syringe pumps to achieve flow throughout the channels.

Using a Formlabs SLA 3D printer, the plates and channels will be fabricated layer by layer, with post-printing processing, including washing and curing. Post-processing will ensure that the device is non-cytotoxic and possesses the transparency needed. Assembly will involve aligning the plates and membrane precisely and securing them together using screws and gasket material. Once the device is assembled, neutrophils will be passed through the top channel, and cytokines will flow through the bottom channel with shear flow. Subsequently, the microfluidic device will be placed on a microscope holder, and fluorescence microscopy will be performed to view the movement of the cells. Some cells will flow straight while others will "duck" through the porous membrane. The microscopy imaging will be analyzed to quantify the movement of the neutrophils and test the efficacy of the device. This methodological approach will enable the investigation of cell behavior under controlled shear stress conditions, providing insights into potential applications for treatments targeting cell migration and interaction.

Results, Conclusions, and Discussions: The fundamental specifications for a new device have been established by the Buffone Lab. In order to facilitate the development of novel pharmacological therapies for blood vessels, the device must functionally allow the investigation of cell migration in an in vitro system. Cytokines should be able to be added to this apparatus by using chemical cues to draw cells across a membrane. Two flow channels are necessary for assembly in order to permit cells to enter the bottom compartment under shear flow conditions or to depart through the outlet. The device has two plates: a top plate and a bottom plate. Laminar flow is achieved on both plates thanks to the input and outlet channel configurations. The two plates are intersected by a single membrane. Both plates shall experience laminar flow as a result of the input and exit channel layouts. Please see the attachments for device specifications. There is just one membrane that crosses over the two plates that will act as the interface between both bottom and top plate. In addition, material requirements include transparency for the purpose of examining cells with fluorescence microscopy, non-cytotoxicity, leak-proofing, and a smooth, flat surface that is essential for preserving laminar flow and undisturbed cells. Lastly, dimensional compatibility is significant, with plates made to easily slide into a microscope holder.
If our device is successful, it can be used to study cell transmigration through a membrane. This technology can help develop diagnostic techniques and improve current drug delivery methods. The development of the microfluidic device as a Point of Care testing device can enable POC testing to be conducted in areas where access to healthcare resources is limited. The devices offer small size, portability, and rapid detection times, making them ideal for POC testing. Particularly in underserved areas lacking trained personnel or laboratory infrastructure, microfluidic POC devices offer a solution to improve access to diagnostic testing.

Acknowledgements (Optional): This work is supported by NIH R01GM143357 to ABJ.