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Tissue Differentiation of ESC into Lung Cells and Functional Validation

  • Ena Ray Banerjee
Chapter

Abstract

The pulmonary system is composed of a variety of epithelial cell populations residing in distinct anatomical locations. Of these, the alveolar epithelial gas exchange surface consists of two cell types, the type I and type II pneumocytes, also known as alveolar epithelial type I and type II (AEI and AEII) cells, that comprise ~95 % and 5 %, respectively, of the alveolar lining area (Chen et al. 2004). AEI cells, important in the regulation of alveolar fluid balance (Dahlin et al. 2004), are branched cells with cytoplasm extremely attenuated for gas exchange (Weibel 1984). AEII cells are cuboidal cells situated between AEI cells and contain characteristic lamellar bodies and apical microvilli (Weibel 1984). Functions of AEII cells include the secretion and reuptake of pulmonary surfactant (Fehrenbach 2001), regulation of alveolar fluid, and synthesis of immunomodulatory proteins (e.g., surfactant protein (SP)-A, SP-D) important for host defense (Matthay et al. 2002). The non-ciliated columnar Clara cells (Evans et al. 1978) constitute the majority of the bronchiolar and terminal bronchiolar epithelium. Clara cells actively divide and differentiate to form ciliated cells, secrete glycosaminoglycans that are major component of the extracellular matrix (ECM), and metabolize airborne toxins by cytochrome P-450 enzymes present in their smooth endoplasmic reticulum (Bishop 2004).

Keywords

Idiopathic Pulmonary Fibrosis Embryoid Body Clara Cell Pluripotent Marker Embryonic Stem Cell Medium 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

AEI

Alveolar epithelial type I

AEII

Alveolar epithelial type II

AQP-5

Aquaporin-5

BAL

Bronchoalveolar lavage

BEGM

Bronchial epithelial growth medium

BFU-E

Burst-forming unit-erythroid

CBP

CREB-binding protein

CC-10

Clara cell-specific protein-10

CFU-E

Colony-forming unit-erythroid

CFU-G

Colony-forming unit-granulocyte

CFU-GEMM

Colony-forming unit-granulocyte/erythroid/macrophage/megakar-yocyte

CFU-GM

Colony-forming unit-granulocyte/macrophage

CFU-M

Colony-forming unit-macrophage

EB

Embryoid body

ECM

Extracellular matrix

FACS

Fluorescence-activated cell sorting

FGF

Fibroblast growth factor

hES

Human embryonic stem

IF

Immunofluorescence

IPF

Idiopathic pulmonary fibrosis

LEF-1

Lymphoid enhancer factor-1

qPCR

Quantitative real-time PCR

SAGM

Small airway growth medium

SP

Surfactant protein

TCF

T cell factor

TGF

Transforming growth factor

VEGF

Vascular endothelial growth factor

Notes

Acknowledgments

We thank Angelique Nelson, Marilyn Nourse, and Carol B. Ware for technical assistance in the culture of hES cells and Bobbie Schneider for technical assistance with the transmission electron microscopy.

Author Contributions

Conceived and designed the experiments: ERB, WRH. Performed the experiments: ERB. Analyzed the data: ERB, WRH. Contributed reagents/materials/analysis tools: MAL, TP, MK, CEM. Wrote the paper: ERB, WRH.

Competing Interests

The authors have declared that no competing interests exist. This does not alter our adherence to all the PLoS ONE policies on sharing data and materials.

Funding

Supported by National Institutes of Health grant R01 HL73722. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Copyright information

© Springer India 2014

Authors and Affiliations

  • Ena Ray Banerjee
    • 1
  1. 1.Department of ZoologyUniversity of CalcuttaKolkataIndia

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