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

PLoS One. 2012;7(12):e50723. doi: 10.1371/journal.pone.0050723. Epub 2012 Dec 7.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipocytes / cytology*
  • Cell Transdifferentiation / physiology
  • Coculture Techniques
  • Hepatocytes / cytology*
  • Humans
  • Spheroids, Cellular / cytology*
  • Stem Cells / cytology*

Grants and funding

This research was supported by the Converging Research Center Program funded by the Ministry of Education, Science and Technology (No. 2012K001360) and the National Research Laboratory (NRL) program, the Korea Science and Engineering Foundation (KOSEF), Republic of Korea (No. 2012026340). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.