Functional 3D human primary hepatocyte spheroids made by co-culturing hepatocytes from partial hepatectomy specimens and human adipose-derived stem cells

• Published in: PloS one Vol. 7; no. 12; p. e50723
• Main Authors: No, Da Yoon, Lee, Seung-A, Choi, Yoon Young, Park, DoYeun, Jang, Ju Yun, Kim, Dong-Sik, Lee, Sang-Hoon
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 07.12.2012; Public Library of Science (PLoS)
• Subjects: Adipocytes - cytology; Albumin; Biology; Biomedical engineering; Biomedical materials; Cell Transdifferentiation - physiology; Coculture Techniques; Cytochrome; Cytochrome P450; Engineering; Enzymatic activity; Hepatectomy; Hepatocytes; Hepatocytes - cytology; Humans; Liver; Liver diseases; Liver transplants; Materials Science; Paracrine signalling; Penicillin; Plastic surgery; Rodents; Secretion; Spheroids; Spheroids, Cellular - cytology; Stem cells; Stem Cells - cytology; Urea; Viability
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0050723

We have generated human hepatocyte spheroids with uniform size and shape by co-culturing 1∶1 mixtures of primary human hepatocytes (hHeps) from partial hepatectomy specimens and human adipose-derived stem cells (hADSCs) in concave microwells. The hADSCs in spheroids could compensate for the low viability and improve the functional maintenance of hHeps. Co-cultured spheroids aggregated and formed compact spheroidal shapes more rapidly, and with a significantly higher viability than mono-cultured spheroids. The liver-specific functions of co-cultured spheroids were greater, although they contained half the number of hepatocytes as mono-cultured spheroids. Albumin secretion by co-cultured spheroids was 10% higher on day 7, whereas urea secretion was similar, compared with mono-cultured spheroids. A quantitative cytochrome P450 assay showed that the enzymatic activity of co-cultured spheroids cultured for 9 days was 28% higher than that of mono-cultured spheroids. These effects may be due to the transdifferentiation potential and paracrine healing effects of hADSCs on hHeps. These co-cultured spheroids may be useful for creating artificial three-dimensional hepatic tissue constructs and for cell therapy with limited numbers of human hepatocytes.