human induced pluripotent stem cells derived airway epithelium

Commonly used acronym: iPSC

Scope of the method

The Method relates to
  • Human health
The Method is situated in
  • Translational - Applied Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Animal derived cells / tissues / organs

Description

Method keywords
  • Human induced Pluripotent Stem Cell
  • differentiation
  • airways
  • lung epithelial cells
Scientific area keywords
  • Induced pluripotent stem cells
  • human airways
  • 3D organoid models
  • Chronic obstructive pulmonary disease
  • asthma
  • epithelial cells
  • co-culture
Method description

We devised a simple and reliable method for reprogramming peripheral blood mononuclear cells into hiPSC and then to differentiate them into air-liquid interface bronchial epithelium (iALI) within 45 days. Of note, this method does not involve any cell sorting step. We reprogrammed blood cells from one healthy control and three patients with very severe COPD. The mean cell purity at definitive endoderm and ventral anterior foregut endoderm was >80%, assessed by CXCR4 and NKX2.1 expression respectively. vAFE cells from all four hiPSC differen-tiated into bronchial epithelium in air-liquid interface (ALI) conditions, with large zones covered by beating ciliated, basal, goblets, club cells and neuroendocrine cells, as found in vivo.

Lab equipment
  • Flow cytometry,
  • Biosafety cabinet,
  • Microscopy phase contrast,
  • Fluorescence PCR,
  • Matrix (geltrex, matrigel),
  • Transwell insert for culture,
  • Transepithelial/transendothelial electrical resistance.
Method status
  • History of use
  • Internally validated
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages
  • Robust, efficient and reproducible protocol
  • Human normal lung development modeling and diseases modeling
  • Stem cells: renewable and sustainable source of airway epithelial cells
  • High-Yield Human Induced Pluripotent Stem Cell-Derived airway epihelial cells
  • Personalized medicine
  • High input pharmacological screening
  • Genome editing CRISPR Cas9 technology
Challenges
  • Cost,
  • Mandatory to check regulary genetic stability of stem cells during culture maintenance,
  • Derivation of clinical grade iPSC culture and derived therapeutic cells.
Modifications
  • Optimization of differentiation protocol,
  • Co-culture with other cell type such as immune cells
Future & Other applications

Disease modeling cell therapy

References, associated documents and other information

References

Differentiation protocol frome the team PMID: 35954266 iPSC cell lines derived PMID: 34624616 , PMID: 33099111, PMID: 30296669 Kotton DN protocol lab: PMID: 35781291, PMID: 35499347 Gotoh Lab: PMID: 34798066

Contact person

Engi Ahmed

Organisations

VIB - UGent
Center for Center for Inflammation Research
Laboratory of Immunoregulation
Belgium
Flemish Region

IRMB INSERM 1183
IRMB INSERM 1183
Belgium
Flemish Region