Guiding Hepatic Differentiation of Pluripotent Stem Cells Using 3D Microfluidic Co-Cultures with Human Hepatocytes
Pouria Fattahi,
Jose M. de Hoyos-Vega,
Jong Hoon Choi,
Caden D. Duffy,
Alan M. Gonzalez-Suarez,
Yuji Ishida,
Kianna M. Nguyen,
Kihak Gwon,
Quinn P. Peterson,
Takeshi Saito,
Gulnaz Stybayeva,
Alexander Revzin
Affiliations
Pouria Fattahi
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Jose M. de Hoyos-Vega
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Jong Hoon Choi
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Caden D. Duffy
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Alan M. Gonzalez-Suarez
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Yuji Ishida
Department of Medicine, Division of Gastrointestinal and Liver Diseases, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA
Kianna M. Nguyen
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Kihak Gwon
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Quinn P. Peterson
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Takeshi Saito
Department of Medicine, Division of Gastrointestinal and Liver Diseases, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA
Gulnaz Stybayeva
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Alexander Revzin
Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA
Human pluripotent stem cells (hPSCs) are capable of unlimited proliferation and can undergo differentiation to give rise to cells and tissues of the three primary germ layers. While directing lineage selection of hPSCs has been an active area of research, improving the efficiency of differentiation remains an important objective. In this study, we describe a two-compartment microfluidic device for co-cultivation of adult human hepatocytes and stem cells. Both cell types were cultured in a 3D or spheroid format. Adult hepatocytes remained highly functional in the microfluidic device over the course of 4 weeks and served as a source of instructive paracrine cues to drive hepatic differentiation of stem cells cultured in the neighboring compartment. The differentiation of stem cells was more pronounced in microfluidic co-cultures compared to a standard hepatic differentiation protocol. In addition to improving stem cell differentiation outcomes, the microfluidic co-culture system described here may be used for parsing signals and mechanisms controlling hepatic cell fate.
