CHARACTERIZATION OF RESPIRATORY CELL SUBPOPULATIONS

Project: Research project

Project Details

Description

In this proposal we attempt to develop experimental approaches for the
isolation, identification and cell culture of tracheal epithelial and
mesenchymal cell subpopulations. The sources of cells will be hamster and
baboon tracheas. We intend to utilize a variety of reagents to
discriminate among and isolate different airway cells using laser flow
cytometry (LFC) and differential bacterial adherence. In conjunction with
multiparameter LFC, the reagents to be used include lectins, and monoclonal
and polyclonal antibodies to cell surface components (markers) such as
fibronectin, keratin and laminin. In one instance, monoclonal antibody and
its complementary bacterial antigen believed to be involved in the
attachment of pathogen (Mycoplasma pneumoniae) to respiratory cells will be
used to separate cells by LFC. The separation of respiratory cells will
also be attempted using fixed, bacterial monolayers of organisms known to
possess binding affinities for eukaryotic cells. Various
immunocytochemical electron microscopic techniques (e.g., Staphylococcus
Protein A-colloidal gold, lectin-colloidal gold) will be used for
ultrastructural examination of freshly isolated and passaged cells to aid
in their identification and assessment of purity. Further long term goals
include the cell culture of isolated or enriched cell populations obtained
by the above methods and their biochemical study in vitro. Recent
successes in the culture of respiratory epithelial cells attest to the
feasibility of such studies. The isolation and biochemical analysis of
specific repiratory cell populations is just in its infancy and may
significantly enhance our understanding of pathological conditions such as
chronic bronchitis, cystic fibrosis and asthma.
StatusFinished
Effective start/end date6/1/8411/30/87

Funding

  • National Institutes of Health
  • National Institutes of Health

ASJC

  • Medicine(all)

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