Microfluidic Genipin Deposition Technique for Extended Culture of Micropatterned Vascular Muscular Thin Films

Eric S. Hald; Kerianne E. Steucke; Jack A. Reeves; Zaw Win; Patrick W. Alford

doi:10.3791/52971

Microfluidic Genipin Deposition Technique for Extended Culture of Micropatterned Vascular Muscula...

JoVE Journal
Bioengineering

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Bioengineering

Microfluidic Genipin Deposition Technique for Extended Culture of Micropatterned Vascular Muscular Thin Films

Full Text

8,150 Views

12:03 min

June 26, 2015

DOI: 10.3791/52971-v

Eric S. Hald¹, Kerianne E. Steucke¹, Jack A. Reeves¹, Zaw Win¹, Patrick W. Alford¹

¹Department of Biomedical Engineering,University of Minnesota

Overview

This study presents a method for microfluidic deposition of genipin and fibronectin on PDMS substrates, enhancing the viability of vascular smooth muscle cell-dense tissues. The approach integrates vascular muscular thin film technology to assess vascular contractility over relevant disease time courses.

Key Study Components

Area of Science

Neuroscience
Vascular Biology
Tissue Engineering

Background

Vascular dysfunctions, such as cerebral vasospasms, are critical health issues.
Existing tissue fabrication methods often lead to rapid degradation.
Long-term culture of vascular tissues is essential for studying contractile functions.
Microfluidic techniques can enhance tissue patterning and viability.

Purpose of Study

To develop an in vitro model for studying smooth muscle contractile function.
To investigate mechanisms underlying vascular dysfunctions.
To maintain tissue integrity and functional contractility over extended periods.

Methods Used

Preparation of elastomer substrates through serial spin coating.
Microfluidic deposition of genipin and fibronectin.
Contractility assays to assess tissue function.
Imaging of samples during contraction and relaxation phases.

Main Results

Tissues maintained structural fidelity and contractility for up to two weeks.
The method outperformed previous techniques in terms of tissue longevity.
Contractility assays provided insights into vascular smooth muscle behavior.
Enhanced tissue organization was observed with the patterned deposition.

Conclusions

This microfluidic method significantly improves the viability of vascular tissues.
It offers a reliable platform for studying vascular contractility over time.
The approach can be applied to investigate various vascular dysfunctions.

Frequently Asked Questions

What is the significance of using PDMS substrates?

PDMS substrates provide a flexible and biocompatible environment for tissue culture, enhancing cell viability.

How does microfluidic deposition improve tissue fabrication?

Microfluidic deposition allows for precise control over the patterning of biomaterials, promoting better tissue organization.

What are the advantages of this method over traditional techniques?

This method maintains tissue integrity and functional contractility for longer periods compared to traditional methods.

Can this model be used to study other vascular conditions?

Yes, the model can be adapted to study various vascular dysfunctions beyond cerebral vasospasms.

What is the role of genipin in this study?

Genipin is used as a crosslinking agent to enhance the mechanical properties of the tissue.

How long can the tissues be maintained in culture?

The tissues can be maintained for up to two weeks while retaining their functional properties.

We present a method for microfluidic deposition of patterned genipin and fibronectin on PDMS substrates, allowing extended viability of vascular smooth muscle cell-dense tissues. This tissue fabrication method is combined with previous vascular muscular thin film technology to measure vascular contractility over disease-relevant time courses.

The overall goal of this procedure is to develop an in vitro model of smooth muscle contractile function that can be used to study the mechanisms of vascular dysfunctions, such as cerebral vasospasms over disease relevant timescales. This is accomplished with long-term vascular muscular thin films by first preparing elastomer substrates by serially spin coating cover slips with a strip of piam followed by PDMS. The second step is to use a microfluidic deposition device to serially deposit genin and fibronectin onto the surface.

In order to provide guidance cues for tissue self-organization on vascular, muscular thin films, the final step is to perform a contractility assay by cussing vascular, muscular thin films inducing contraction and relaxing contraction while imaging the sample. Ultimately, this approach yields tissues that maintain structural fidelity and functional contractility for up to two weeks in culture. The main advantage of this technique over existing methods like micro contact printed muscular thin films, is that tissue integrity and functional contractility are maintained for weeks where tissues constructed using previous methods begin to degrade after three to four days.

View the full transcript and gain access to thousands of scientific videos