Hepatic Organoid Formation in Collagen Sponge of Cells Isolated from Human Liver Tissues

Abstract

We examined whether small hepatocytes (SHs), which are hepatic progenitor cells, could be isolated from a normal human liver and whether human hepatic cells could form hepatic organoids in a collagen sponge. Normal liver tissues were obtained from resected specimens from nine patients who underwent hepatic resection. Isolated hepatic cells were plated on dishes and a collagen sponge. More than 1 month later, SH-like cells appeared and proliferated on the dishes, whereas cell aggregates were formed in the sponge and showed characteristic tissue architecture: columnar and/or cuboidal epithelial cells lined the surface of the sponge. Clusters of epithelial cells with a large cytoplasm and ductular structures were observed under the lining cells. The lining and ductular cells were positive for cytokeratins 7 and 19, which indicated they were biliary epithelial cells (BECs), and the epithelial cells forming clusters were positive for the anti-human hepatocyte antibody, identifying them as hepatocytes. Some lining cells were positive for both the hepatic marker and the BEC markers. The cells in the collagen sponge actively proliferated and the hepatocytes excreted albumin into the medium. Thus, hepatic organoids could be reconstructed in a collagen sponge by normal human liver cells.

The science and engineering of stem cell‐derived organoids‐examples from hepatic, biliary, and pancreatic tissues

“…Compared to the use of mature parenchymal cells that are terminally differentiated, hepatoblast‐like hepatic progenitor cells have the potential to divide and differentiate, and may self‐organize better into a 3D liver‐like tissue. In one of the first studies of hHOs, donor (adult) human small hepatocytes (SHs) and non‐parenchymal liver cells, cultured in an organoid format within collagen sponges processed from bovine tendon, produced functional hepatocyte‐like cells that were spatially segregated to the outer layer, some biliary duct formation, and surrounding vascular structures (Sugimoto et al ., 2005). This study demonstrates several principles of organoids, including the use of progenitors, spatial segregation and aggregation, morphogenesis, and differentiation.…”

Cryo-chemical decellularization of the whole liver for mesenchymal stem cells-based functional hepatic tissue engineering

“…polylactic acid, polyglycolic acid, and polyethylene glycol hydrogels) [8–10] and natural (e.g. collagen, alginate and chitosan) [11–14] three-dimensional (3D) scaffolds that provide an environment supporting the maintenance and growth of hepatocytes. More recently, studies have shown that organ decellularization is an attractive strategy because it allows the creation of a naturally occurring 3D biomimetic scaffold that is available for tissue engineering [15,16].…”

Concise Review: The Relevance of Human Stem Cell-Derived Organoid Models for Epithelial Translational Medicine

“…There are currently two main approaches for generating physiologically relevant liver organoids: sandwich cultures and self-assembling spheroids 28, 31 . In sandwich culture models, patient-derived hepatocyte progenitor cells are seeded between layers of collagen or Matrigel in order to replicate in vivo liver morphology and maintain epithelial hepatocyte polarity 28 .…”

“…In sandwich culture models, patient-derived hepatocyte progenitor cells are seeded between layers of collagen or Matrigel in order to replicate in vivo liver morphology and maintain epithelial hepatocyte polarity 28 . In contrast, self-assembling spheroids are formed when hepatocyte progenitor cells are cultured alongside stromal cells in a collagen sponge 31, 32 . Spheroids such as these more closely resemble in vivo hepatocyte physiology with cells forming better tight junctions, cell polarity, protein expression and metabolic activity.…”