Page 7 - ebook

The Ciliary Movement and Maturation on the Ciliated Cell with Endometriosis
1
2
Seung Hee Kim , Joo Hyeon Park , JinAh Lee , Young Min Shin and Jung Bok Lee *
1
1,
1
1 Department of Biological Science, Sookmyung Women’s University, Seoul 04310, Korea
2 Department of Medical Engineering, Yonsei University College of Medicine, Seoul 03722, Korea
*E-mail : jungboklee@sookmyung.ac.kr
INTRODUCTION RESULTS & DISCUSSION
• Endometriosis is estrogen-dependent disorder • The mRNA expression levels in the epithelial
defined by presence and growth of endometrial cells
tissue outside the uterus. 1.5 DNAI1 DNALI1

• We investigated the correlation of endometriosis 1.0 * 1.5 *
1.0
and ciliary movement or maturation of ciliated cell. Relative mRNA transcripts 0.5 Relative mRNA transcripts 0.5

• Human primary epithelial cells from the normal 0.0 0.0 * EMS: endometriosis
and endometriosis fallopian tube were cultured on Normal P6 EMS P6 Normal P6 EMS P6
a microfluidic chip with a 500 µm width. Different • RT-qPCR quantification of mRNA expression to compare cilia
flow rates are generated on the microchannel to motor protein from normal and endometriosis fallopian tube
generate shear stress for produce and maturation of epithelial cells, as presented by marker expression of the outer
dynein arm (DNAI1) and the inner dynein arm (DNALI1).
cilia on the ciliated cell.
Fallopian tube epithelial cell • Comparison of Acetylated tubulin expression
levels

Ciliated cell Secretory cell

Endometriosis
Created with BioRender.com
* EMS: endometriosis
MATERIALS & METHODS
• Confocal images of normal and endometriosis ciliated cell co-
⚫ Cilia Motor Protein immunostaining of Acetylated tubulin (green) with nucleus
Cilium cross-section (blue). Scale bar indicated 20 µm.
Outer dynein arm • Result of Western blotting on normal and endometriosis
- Dynein Axonemal Intermediate
Chain 1 (DNAI1) epithelial cells. Quantitative analysis of the expression in
comparison to GAPDH, an internal control.
Inner dynein arm • Fallopian tube epithelial cell culture under
- Dynein Axonemal Light Intermediate
Chain 1 (DNALI1) flow condition

Poprzeczko, M. et al., Cells 2019;8(12), 1614
• We focused on the correlation of ciliary activity and
pathogenesis of endometriosis.
• Dynein Axonemal Intermediate Chain 1 (DNAI1) and Dynein
Axonemal Light Intermediate Chain 1 (DNALI1) were chosen
as the marker because they are known as cilia motor protein.
• Images of normal and endometriosis fallopian tube epithelial
⚫ Isolation primary cell from normal and endometriosis cells that cultured in the 600 µm width I type slide chip with one
fallopian tube direction constant flow and recorded at day 1, day 5 and day 6.
Scale bar indicated 200 µm.

ONGOING WORK
Currently, we are working towards to fabricate Fallopian tube
chip imitates biological characteristics of fallopian tube and
Created with BioRender.com generate different flow rates on the microchannel to generate
• Fallopian tube specimens were obtained from patients shear stress for produce and maturation of cilia on the ciliated
with/without endometriosis. cell.

CONCLUSION

This system replicates the epithelium of normal and endometriosis to apply to studies of the cause of the endometriosis,
suggesting a promising alternative to animal models.