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2008 Annual Meeting Abstracts

Novel Therapeutic Approaches to Premature Lung Disease Utilizing Stem Cells

Christine Finck, MD1,2, Blair Roszell1, Ariel Seaton1, Guo-Hua Fong, PhD2.
1Connecticut Children's Medical Center, Hartford, CT, USA, 2University of Connecticut Health Center, Hartford, CT, USA.

Objective: Each week in Connecticut, 81 babies are born prematurely. These preterm newborns have pulmonary hypoplasia(PH) with a mortality as high as 91%. PH is characterized by poorly differentiated alveolar epithelium, reduced alveolar type 2 cells (AE2) and decreased surfactant production. Recently we demonstrated a method for deriving AE2-like cells from pluripotent murine embryonic stem cells (mESC). We hypothesize that these differentiated cells can be intra-tracheally administered into a mouse and incorporate into the recipient lung parenchyma. This represents a first step toward stem cell based treatment of PH.
Design: CD4-Foxa2/GFP-Bry murine ESCs (mESC) were cultured in Activin A and switched to media containing bFGF, yielding 12.4% surfactant protein C (SpC) expressing cells(AE2). These cells were suspended in media (100,000 cells/ml). Adult 129 mice were anesthetized and 0.1ml of cells or saline (control) was injected intratracheally through a 27 guage needle. The lungs from the recipient mice were harvested and fixed after 7 days.
Outcome Measures: Immunohistochemical identification of lung intra-parenchymal mESC-derived AE2 cells using antibodies for SpC (red) and CD4 (green) with nuclear BBZ counterstaining. Double staining (yellow) identifies intratracheally administered mESC derived AE2 cells.
Results: Incorporation of ESC-derived AE2 cells within the host lung parenchyma (arrow) was evident. The cells were located in the distal alveoli and did not aggregate in the proximal airways.
Conclusions: We demonstrate that delivery of mESC derived AE2 cells is feasible with intratracheal injection. This represents the first step towards translational research with site-specific delivery for a stem cell-based therapeutic approach towards PH.


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