Novel Methods for Delivery of Cell-Based Therapies

Cecilia Crisanti, M.; Koutzaki, Sirma H.; Mondrinos, Mark J.; Lelkes, Peter I.; Finck, Christine M.

doi:10.1016/j.jss.2007.06.010

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Volume 146, Issue 1 , Pages 3-10, 1 May 2008

Novel Methods for Delivery of Cell-Based Therapies

M. Cecilia Crisanti, M.D.
- Department of Pediatric Surgery, St. Christopher’s Hospital for Children, Drexel University College of Medicine, Philadelphia, Pennsylvania
- Department of Thoracic Oncology, University of Pennsylvania, Philadelphia, Pennsylvania
- To whom correspondence and reprint requests should be addressed at Abramson Research Center, University of Pennsylvania, School of Medicine, 3615 Civic Center Blvd., Suite 1015F, Philadelphia, PA 19104.
Sirma H. Koutzaki, M.S.
- Department of Pediatric Surgery, St. Christopher’s Hospital for Children, Drexel University College of Medicine, Philadelphia, Pennsylvania
Mark J. Mondrinos, B.S.
- School of Biomedical Engineering, Science and Health Systems, Drexel University, Philadelphia, Pennsylvania
Peter I. Lelkes, Ph.D.
- School of Biomedical Engineering, Science and Health Systems, Drexel University, Philadelphia, Pennsylvania
Christine M. Finck, M.D.
- Department of Pediatric Surgery, St. Christopher’s Hospital for Children, Drexel University College of Medicine, Philadelphia, Pennsylvania
- Department of Pediatric Surgery, Connecticut Children’s Hospital, Hartford, Connecticut

Received 8 January 2007 published online 28 July 2007.

Background

Pulmonary hypoplasia (PH) is found in 15% to 20% of all neonatal autopsies, accounting for 2850 deaths yearly. Development of engineered tissue substitutes that could functionally restore damaged tissue remains a unique opportunity for biotechnology. Recently, we isolated and characterized murine fetal pulmonary cells (FPC) and engineered 3-D pulmonary tissue constructs in vitro. Our goal is to devise a reliable and reproducible method for delivering FPC into a live animal model of PH.

Materials and methods

Three methods of delivery were explored: intraoral, intratracheal, and intrapulmonary injection. Adult Swiss Webster mice were anesthetized and fluorescent labeled microspheres (20 μm diameter) were delivered by intraoral and intratracheal injection. Subsequently, labeled FPC (Cell Tracker, CMTPX; Molecular Probes, Eugene, OR) were delivered by the same methods. In addition, direct transpleural intrapulmonary injection of FPC was performed. Outcome analysis included survival, reproducibility, diffuse versus confined location of the injected substance, and adequacy of delivery. Routine histological examination, fluorescent microscopy, and immunostaining were performed.

Results

Microspheres: We demonstrated reproducible, diffuse instillation via tracheotomy into the distal alveoli. Intraoral delivery appeared less reliable compared to direct intratracheal injection. FPC: Intratracheal injection was a reliable method of delivery. Labeled FPC showed transepithelial migration after 7 d of in vivo culture. Intrapulmonary injection led to local accumulation of cells in sites of injection.

Conclusions

We demonstrate that delivery of FPC is feasible with intratracheal injection giving the most reliable, diffuse delivery throughout the lung. This represents the first step toward translational research with site-specific delivery for a cell-based therapeutic approach toward PH and similar pulmonary diseases.

Key Words: cell-based therapy, pulmonary hypoplasia, murine fetal pulmonary cells, intratracheal injection, intraoral injection, intrapulmonary injection, transepithelial migration