DECIPHERING MSC DIVERSITY AND INTERACTION WITH EPITHELIA
John DE VOS
Tel: + 33 4 67 33 57 23
Email : email@example.com
DE VOS John (PUPH/UM)
ASSOU Said (IR/UM)
NASRI Amel (PhD-Student)
FOISSET Florent (PhD-Student)
MIANNE Joffrey (PhD-student)
FIELDES Mathieu (Ph-D-student)
BOURDAIS Carine (PhD-Student)
Pluripotent stem cell
Primary ciliary dyskinesia
Primary peripheral blood mononuclear cells (PBMC) procurement; cell reprogramming; pluripotent stem cell culture; differentiation of iPSC into bronchial epithelium; transepithelialelectrical resistance (TEER)
mRNAseq, single cell mRNAseq, functional bioinformatics
RT-qPCR, ddPCR copy number counting
Imaging, live video:
Confocal microscopy; scanning and transmission electronmicroscopy (SEM and TEM); high-speed camera capture and computing of cilia beating frequency and orientation.
Epithelia, that line the outer surfaces of organs and blood vessels throughout the body, as well as the inner surfaces of cavities in many internal organs, have critical functions in the body, including protection, secretion and exchange. Their development and function rely on a close connection with underlying connective tissue, which contains MSC. Thus, MSC play a crucial role in epithelia, an often overlooked role of these cells.
In close collaboration with the team of Pr. Arnaud Bourdin (U1048), we developed a cutting edge model of airway development in vitro, using induced pluripotent stem (iPS) cells obtained from peripheral blood mononuclear cells (PBMC) from healthy subjects and from patients with chronic inflammatory lung disease, i.e. COPD. After comprehensive benchmarking, we developed a method to recapitulate the sequential processes that progressively drive pluripotent stem cells into endoderm (DE), proximal lung progenitors (vAFE) and then proximal lineage-specific bronchial epithelial cells (BEC) (Ahmed PT 2018, Ahmed et al, submitted). This 2D bronchial epithelium shows all the characteristic features of a functional airway epithelium as occurs in vivo with presence of ciliated, goblet, basal and club cells; additionally, transepithelial electrical resistance (TEER) can be successfully acquired. This bronchial epithelial develops on a mesenchymal layer, which role is uncertain, providing a platform to evaluate the role of the mesenchymal compartment on the epithelium development and turn-over. Moreover, leveraging on CRISPR/Cas9 technology, we are repairing iPS cell lines derived from peripheral blood mononuclear cells from patient with primary ciliary dyskinesia (PCD) to restore in vitro cilia movement.