Comparative study between lactose-silk fibroin conjugates and extracellular matrices as a substrate for the culture of human induced pluripotent stem cell-derived hepatocytes
Affiliations: [a] Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 1-2 Ohwashi, Tsukuba, Ibaraki, Japan | [b] Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kanagawa, Japan | [c] REPROCELL Inc., 3-8-11 Shin-yokohama, Kohoku-ku, Yokohama, Kanagawa, Japan
Abstract: BACKGROUND:Human induced pluripotent stem cell (hiPSC)-derived hepatocytes are an attractive alternative cell source to primary human hepatocytes for tissue regeneration. OBJECTIVES:This study presents an application of lactose-silk fibroin conjugates (Lac-CY-SF) bearing 𝛽-galactose residues as a substrate for culture of hiPSC-derived hepatocytes. A comparison of hiPSC-derived hepatocytes cultured on three different substrates; Lac-CY-SF conjugates, Matrigel and type I collagen was performed. METHODS:Cell morphology, viability, maturation and albumin secretory function were assessed by phase-contrast microscopy, tetrazolium-based colorimetric assay, immunofluorescence staining and enzyme-linked immunosorbent assay. RESULTS:Morphological characteristics of the cells cultured on the conjugates resembled those on Matrigel throughout the 6-day culture period. The number of viable cells cultured on the conjugates was comparable to that on Matrigel at day 2 and 6. The protein expression of mature hepatocyte markers, asialoglycoprotein receptor 1 and albumin, by the cells cultured on the conjugates resembled that by the cells cultured on collagen at day 2 and 6. Albumin secretory function per cell cultured on the conjugates was higher than that on collagen and comparable to that on Matrigel. CONCLUSIONS:These limited results suggest that Lac-CY-SF conjugates may be as useful as Matrigel and collagen for cultivation of hiPSC-derived hepatocytes.
