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1 Centre for Early Human Development, Monash Institute of Medical Research, Melbourne, Victoria, Australia
2 Department of Biochemsitry and Molecular Biology, Monash University, Melbourne, Victoria, Australia
3 Centre for Early Human Development, Monash Institute of Medical Research, Melbourne, Victoria, Australia; Department of Biochemsitry and Molecular Biology, Monash University, Melbourne, Victoria, Australia
* To whom correspondence should be addressed. E-mail: Richard.Mollard{at}med.monash.edu.au.
Mouse embryonic stem cells (MESCs) are pluripotent, theoretically immortal cells derived from the inner cell mass of developing blastocysts. The respiratory epithelium develops from the primitive foregut endoderm as a result of inductive morphogenetic interactions with the surrounding visceral mesoderm. Following dissociation of the explanted fetal lung into single cells, these morphogenetic signalling pathways instruct reconstitution of the developing lung according to a process known as organotypic regeneration. Data presented here demonstrate that such fetal lung morphogenetic cues induce MESC derivatives to incorporate into the reforming pseudoglandular-like tubular ducts, display pan-keratin and surfactant protein C (Sftpc) immunoreactivity and express Sftpc transcripts while displaying a normal diploid karyotype in co-culture. The Sftpc inductive capacity of dissociated fetal lung tissue shows stage specificity with 24% of all MESC derivatives displaying Sftpc immunoreactivity following co-culture with embryonic day 11.5 (E11.5) lung buds, compared with 6% and 0.02% following co-culture with E12.5 and E13.5 lung buds, respectively. MESC derivative Sftpc immunoreactivity follows a spatial and temporal specific maturation profile with an initially ubiquitous cellular Sftpc immunostaining pattern becoming apically polarised with time. Directing differentiation of MESCs into respiratory lineages has important implications for cell replacement therapeutics aimed at treating respiratory-specific diseases such as cystic fibrosis and idiopathic pulmonary fibrosis.
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