Transplantation of patient-specific bile duct bioengineered with chemically reprogrammed and microtopographically differentiated cells

Elina Maria Buisson; Suk-Hee Park; Myounghoi Kim; Kyojin Kang; Sangtae Yoon; Ji Eun Lee; Young Won Kim; Nak Kyu Lee; Mi Ae Jeong; Bo-Kyeong Kang; Seung Bum Lee; Valentina M Factor; Daekwan Seo; Hyunsung Kim; Jaemin Jeong; Han Joon Kim; Dongho Choi

doi:10.1002/btm2.10252

Transplantation of patient-specific bile duct bioengineered with chemically reprogrammed and microtopographically differentiated cells

Bioeng Transl Med. 2021 Sep 3;7(1):e10252. doi: 10.1002/btm2.10252. eCollection 2022 Jan.

Authors

Elina Maria Buisson^{1

2}, Suk-Hee Park³, Myounghoi Kim^{1

2}, Kyojin Kang^{1

2}, Sangtae Yoon^{1

2}, Ji Eun Lee⁴, Young Won Kim^{4

5}, Nak Kyu Lee⁴, Mi Ae Jeong⁶, Bo-Kyeong Kang⁷, Seung Bum Lee⁸, Valentina M Factor⁹, Daekwan Seo¹⁰, Hyunsung Kim¹¹, Jaemin Jeong^{1

2}, Han Joon Kim^{1

2}, Dongho Choi^{1

2}

Affiliations

¹ Department of Surgery Hanyang University College of Medicine Seoul Republic of Korea.
² HY Indang Center of Regenerative Medicine and Stem Cell Research Hanyang University Seoul Republic of Korea.
³ School of Mechanical Engineering Pusan National University Busan Republic of Korea.
⁴ Digital Manufacturing Process Group Korea Institute of Industrial Technology Siheungsi Gyeonggi-do Republic of Korea.
⁵ Present address: Current address: School of Mechanical Engineering Purdue University West Lafayette Indiana USA.
⁶ Department of Anesthesiology and pain medicine Hanyang University College of Medicine Seoul Republic of Korea.
⁷ Department of Radiology Hanyang University, College of medicine Seoul Republic of Korea.
⁸ Laboratory of Radiation Exposure & Therapeutics National Radiation Emergency Medical Center, Korea Institute of Radiological & Medical Science Seoul Republic of Korea.
⁹ Laboratory of Molecular Pharmacology Center for Cancer Research, National Cancer Institute, National Institutes of Health Bethesda Maryland USA.
¹⁰ Psomagen Inc Rockville Maryland USA.
¹¹ Department of Pathology Hanyang University College of Medicine Seoul Republic of Korea.

Abstract

Cholangiopathy is a diverse spectrum of chronic progressive bile duct disorders with limited treatment options and dismal outcomes. Scaffold- and stem cell-based tissue engineering technologies hold great promise for reconstructive surgery and tissue repair. Here, we report a combined application of 3D scaffold fabrication and reprogramming of patient-specific human hepatocytes to produce implantable artificial tissues that imitate the mechanical and biological properties of native bile ducts. The human chemically derived hepatic progenitor cells (hCdHs) were generated using two small molecules A83-01 and CHIR99021 and seeded inside the tubular scaffold engineered as a synergistic combination of two layers. The inner electrospun fibrous layer was made of nanoscale-macroscale polycaprolactone fibers acting to promote the hCdHs attachment and differentiation, while the outer microporous foam layer served to increase mechanical stability. The two layers of fiber and foam were fused robustly together thus creating coordinated mechanical flexibility to exclude any possible breaking during surgery. The gene expression profiling and histochemical assessment confirmed that hCdHs acquired the biliary epithelial phenotype and filled the entire surface of the fibrous matrix after 2 weeks of growth in the cholangiocyte differentiation medium in vitro. The fabricated construct replaced the macroscopic part of the common bile duct (CBD) and re-stored the bile flow in a rabbit model of acute CBD injury. Animals that received the acellular scaffolds did not survive after the replacement surgery. Thus, the artificial bile duct constructs populated with patient-specific hepatic progenitor cells could provide a scalable and compatible platform for treating bile duct diseases.

Keywords: 3D printing; chemically derived hepatic progenitors; cholangiocytes; electrospun fibers; patient‐specific bile duct.