Kidney Week

Abstract: TH-OR036

Kidney Proximal Tubule Engineering via Melt-Electrowriting

Session Information

• Bioengineering
  November 07, 2019 | Location: 146 A/B, Walter E. Washington Convention Center
  Abstract Time: 05:30 PM - 05:42 PM

Category: Bioengineering

• 300 Bioengineering

Authors

• Jansen, Katja, Utrecht Institute for Pharmaceutical Sciences, Utrecht, Netherlands

Katja Jansen



Katja Jansen obtained her MSc in Biomedical Sciences at Radboud University, Nijmegen, The Netherlands. Currently, she works as PhD candidate at Utrecht University, The Netherlands, under supervision of Prof. Roos Masereeuw. Her aim is the development of advanced therapies for kidney patients. She focuses on the use of proximal tubule cells for the fabrication of kidney tubes, and studies pathways involved in transporter regulation.

• Genderen, Anne metje Van, Utrecht University, Utrecht, Netherlands

Anne metje Van Genderen,

• Kristen, Marleen, Utrecht University, Utrecht, Netherlands

Marleen Kristen,

• van Duijn, Joost, University Medical Center Utrecht, Utrecht, Netherlands

Joost van Duijn,

• Vermonden, Tina, Utrecht University, Utrecht, Netherlands

Tina Vermonden,

• Malda, Jos, UMC Utrecht, Utrecht, Netherlands

Jos Malda,

• Masereeuw, Rosalinde, Utrecht Institute for Pharmaceutical Sciences, Utrecht, Netherlands

Rosalinde Masereeuw,

• Castilho, Miguel, UMC Utrecht, Utrecht, Netherlands

Miguel Castilho,

Background

Tubular tissue engineering generally relies on large scaffolds (>1 mm) or smaller tubes within bulk hydrogels. However, for the engineering of kidney tubuli, these structures must preferably be both small-sized to increase surface area and freely accessible for rapid waste removal. Here, we report on the fabrication of a small scale and self-standing living proximal tubule by combining well-organized tubular fiber scaffolds obtained by melt-electrowriting (MEW) with human kidney cells.

Methods

A custom-built MEW device was used to fabricate poly(ε-caprolactone) scaffolds with defined microarchitectures (square, rhombus or random) and inner Øs of 0.5-1 mm (Fig. 1A). Well-characterized human conditionally immortalized proximal tubular epithelial cells (ciPTEC) were seeded inside the scaffolds and tested for monolayer integrity, organization, matrix production and cell functionality.

Results

Scaffolds were manufactured by controlling acceleration voltage, dispensing pressure and mandrel rotation and translation speed. The pore resolution was 200-500 µm (Ø 5-10 µm microfibers, 400 µm scaffold thickness and inner Ø 0.5-1 mm) (Fig. 1B). ciPTEC formed tight monolayers in all scaffolds; rhombus shaped pores facilitated unidirectional cell orientation (Fig. 1C). The cells deposited extracellular matrix (ECM) directly after seeding, and collagen IV quantity increased over time. Viability was proven by enzymatic conversion of calcein-AM into calcein, and active uptake of RH-123; transport inhibitor sensitivity proved cell functionality.

Conclusion

Here, we present self-standing, small diameter kidney tubes that show cell functionality. Due to the well-organized tubular scaffold microstructure with large, interconnected pores, the self-produced ECM is the only barrier between the inner and outer compartment, facilitating rapid and active solute uptake. We are currently evaluating the potential for application in micro-physiological test systems and for fine-tuning towards implantable tissues with sufficient mechanical stability.

Funding

• Government Support - Non-U.S.