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Microencapsulation of Leydig cells

Microencapsulation of Leydig cells. Team: Bryan Baxter Tim Eng Joe Zechlinski April Zehm BME 400 October 14, 2005. Client: Dr. Craig Atwood Dr. Sivan Vadakkadath Meethal Miguel Gallego VA Hospital Advisor: Assistant Professor Kristyn Masters Department of Biomedical Engineering.

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Microencapsulation of Leydig cells

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  1. Microencapsulation of Leydig cells Team: Bryan Baxter Tim Eng Joe Zechlinski April Zehm BME 400 October 14, 2005

  2. Client:Dr. Craig AtwoodDr. Sivan Vadakkadath MeethalMiguel GallegoVA HospitalAdvisor:Assistant Professor Kristyn MastersDepartment of Biomedical Engineering

  3. Overview • Problem Statement • Background • Design Specifications • Recap of previous work • New directions

  4. Problem Statement • Develop method of encapsulating cells to allow hormone release while providing a physical barrier to the host’s immune system Motivation • Potential alternative to less desirable treatments • Organ transplant • Hormone injections • Cellular grafts

  5. Microcapsules • Provide physical barrier to immune system • Consist of hydrogels • Implanted in vivo • Time-released hormone therapy (Uludag et al., 2000)

  6. Client Research • Microencapsulation applications • Anti-aging therapy • Reproductive disorders • Cells and hormones of interest • Leydig and Sertoli cells • Testosterone, inhibin, activin, FSH, LH (Ownby, 1999) (Adapted from Morohashi, 1997)

  7. Biocompatibility Material properties Crosslinking procedure Repeatability of results Immunoprotection Controlled pore size (MWCO) Degradation Mechanical Biological Design Specifications

  8. Previous work • Polyethylene glycol (PEG) • Diacrylated synthetic polymer • UV-crosslinked • Water-in-oil emulsification

  9. Previous work • Bioprinter • Modified Epson R200 inkjet printer • Piezoelectric droplet generation • Linux platform, software interface

  10. Microsphere Production • Microfluidic devices • Increased precision and control on microscale • Sample and sheath flow rates determine size and quantity of microcapsules • Minimal reagents used (Jeong et al., 2005)

  11. Cell culture • Cell line • MA-10 cells (mouse cancer Leydig cells) • Produce testosterone • Cell passaging • Split proliferating cells • Hemocytometry • Cryofreezing (Saltzman, 2004.)

  12. PEG hydrogels on the macroscale • Current Experiments: • Acrylation procedure • Hydrogel swelling/PEG crosslinking • Planned Experiments: • Combine with MA-10 cell suspension 12.5x

  13. (http://respiratory-research.com) Viability/Hormone studies • Live/Dead® assay • Metabolism (greenlive) • Membrane integrity (reddead) • Testosterone assay • Competitive sandwich ELISA (ALPCO Diagnostics)

  14. References Jeong, W., et al. 2005. Continuous fabrication of biocatalyst immobilized microparticles using photopolymerization and immiscible liquids in microfluidic systems. Langmuir 21: 3738-3741. Machluf M, Orsola A, Boorjian S, Kershen R, and Atala A. 2003. Microencapsulation of leydig cells: a system for testosterone supplementation. Endocrinology 144:4975-4979. Morohashi, K. 1997. The ontogenesis of the steroidogenic tissues. Genes to Cells 2: 95-106. Ownby. 1999. Histology: male reproductive system. http://www.cvm.okstate.edu/instruction/mm_curr/histology/MR/HiMRp3.htm. Accessed February 12, 2005. Saltzman, W. 2004. Tissue engineering : engineering principles for the design of replacement organs and tissues. New York : Oxford University Press. Uludag H, De Vos P, and Tresco PA. 2000. Technology of mammalian cell encapsulation. Advanced Drug Delivery Reviews 42:29-64. http://chemfinder.cambridgesoft.com http://respiratory-research.com http://web.indstate.edu/thcme/mwking/glycans.html http://www.cybercolloids.net/library/alginate/structure.php

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