Micropatterned organoid culture of rat hepatocytes and HepG2 cells

Mori, Ryuhei; Sakai, Yusuke; Nakazawa, Kimitaka

doi:10.1263/jbb.106.237

Cited by 46 publications

(29 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To solve this problem, we have three options: (i) selecting several types of cells and organizing them into tissue structure, (ii) selecting the precursor cells for histogenesis of liver, inducing differentiation and prompting self-organization and (iii) constructing artificial culture microenvironments mimicking native environments of liver. The culture microenvironments, such as microwell architecture (Mori et al, 2008), are well known to improve the function of cells, including hepatocytes (Sharma et al, 2010). Previously, we showed the induction of retinal cells by co-culturing with ESC-derived RPE cells (Yue et al, 2010).…”

Section: In Vitro Tissue Formation Using Induced Hepatocytesmentioning

confidence: 99%

A Strategy Using Pluripotent Stem Cell-Derived Hepatocytes for Stem Cell-Based Therapies

Tomotsune¹,

Sato²,

Yoshie³

et al. 2011

Advances in Regenerative Medicine

View full text Add to dashboard Cite

Section: In Vitro Tissue Formation Using Induced Hepatocytesmentioning

confidence: 99%

A Strategy Using Pluripotent Stem Cell-Derived Hepatocytes for Stem Cell-Based Therapies

Tomotsune¹,

Sato²,

Yoshie³

et al. 2011

Advances in Regenerative Medicine

View full text Add to dashboard Cite

“…The remaining part of each microwell was modified with PEG and represented the nonadhesive area. This chip was prepared using the microfabrication and microcontact-printing techniques [Fukuda et al, 2006;Mori et al, 2008]. The chip was immersed in 50% ethanol for sterilization and then thoroughly rinsed with distilled water.…”

Section: Hepatocyte Spheroid Chipmentioning

confidence: 99%

“…Otsuka et al [2004] have developed a multiarray of hepatocyte spheroids on a micropatterned polyethylene glycol (PEG)-brash surface. We have developed a spheroid production technique that involves the use of a microfabricated chip with cylindrical microwells; the bottom surface of each microwell is micropatterned with collagen and PEG [Fukuda et al, 2006;Mori et al, 2008]. These microchip techniques are advantageous over previous methods involving the use of spheroids because they enable mass production of uniform spheroids (or cell aggregates).…”

Section: Introductionmentioning

confidence: 99%

Comparative Analysis of Gene Expression in Rat Liver Tissue and Monolayer- and Spheroid-Cultured Hepatocytes

Sakai

Yamagami

Nakazawa³

2009

Cells Tissues Organs

Self Cite

View full text Add to dashboard Cite

A culture system with spherical multicellular aggregates (spheroids), which are formed by the rearrangement and compaction of cell aggregates, is reported to be more useful than the traditional monolayer culture system for the culture of primary hepatocytes. By performing real-time polymerase chain reaction, we analyzed the expression of genes encoding key molecules involved in liver-specific functions, namely, cell adhesion molecules (integrin 3, cadherin 1 and connexin 32), transcription factors (hepatic nuclear factor 4α and CCAAT/enhancer-binding protein β), protein and metabolic enzymes (albumin, glucose-6-phosphatase, tryptophan 2,3-dioxygenase, arginase 1 and cytochrome P450 7A1) and transporters (organic anion transporting peptide 1, multidrug resistance-associated protein 2 and bile salt export pump), in spheroids derived from rat hepatocytes. Further, we compared these expression levels with those in a hepatocyte monolayer and in liver tissue. Only the gene encoding glucose-6-phosphatase (required for sugar metabolism) was expressed at a similar level in both the monolayer culture and liver tissue for 10 days of culture; the expression of all the other genes in the monolayer culture either rapidly decreased or completely disappeared as the culture duration increased. Although the expression levels of all the genes in the spheroids tended to decrease gradually with culture time, they were consistently higher than those in the monolayer culture for at least 10 days of culture. These results suggest that hepatocyte spheroids acquire intercellular organization and largely maintain many intercellular metabolic functions. Thus, the hepatocyte spheroid culture system seems to be promising for various in vitro cell-based assays.

show abstract

“…Such a 3D culture model has occurred to recapitulate many in vivo tissue structures and functions [3,9]. Very few hepatocyte spheroid models were established using: a poly-(L-lactic acid ) polymer [10], rock techniques [11], micro-rotation flows [12], alginate scaffolds [13], RGD and galactose-conjugated membranes [14], positive-charged substrates [4], micropatterning techniques [15], nanopillar sheets [16], galactosylated nanofiber scaffold [17], or polyurethane forms [18]. However, hepatocyte spheroids under the influence of fetal calf serum and nonparechyalmal cells have not yet been established.…”

Section: Introductionmentioning

confidence: 99%

Morphological and Functional Analysis of Hepatocyte Spheroids Generated on Poly-HEMA-Treated Surfaces under the Influence of Fetal Calf Serum and Nonparenchymal Cells

Açıkgöz

Giri²,

Cho

et al. 2013

Biomolecules

View full text Add to dashboard Cite

Poly (2-hydroxyethyl methacrylate) (HEMA) has been used as a clinical material, in the form of a soft hydrogel, for various surgical procedures, including endovascular surgery of liver. It is a clear liquid compound and, as a soft, flexible, water-absorbing material, has been used to make soft contact lenses from small, concave, spinning molds. Primary rat hepatocyte spheroids were created on a poly-HEMA-coated surface with the intention of inducing hepatic tissue formation and improving liver functions. We investigated spheroid formation of primary adult rat hepatocyte cells and characterized hepatic-specific functions under the special influence of fetal calf serum (FCS) and nonparencymal cells (NPC) up to six days in different culture systems (e.g., hepatocytes + FCS, hepatocytes – FCS, NPC + FCS, NPC – FCS, co-culture + FCS, co-culture – FCS) in both the spheroid model and sandwich model. Immunohistologically, we detected gap junctions, Ito cell/Kupffer cells, sinusoidal endothelial cells and an extracellular matrix in the spheroid model. FCS has no positive effect in the sandwich model, but has a negative effect in the spheroid model on albumin production, and no influence in urea production in either model. We found more cell viability in smaller diameter spheroids than larger ones by using the apoptosis test. Furthermore, there is no positive influence of the serum or NPC on spheroid formation, suggesting that it may only depend on the physical condition of the culture system. Since the sandwich culture has been considered a “gold standard” in vitro culture model, the hepatocyte spheroids generated on the poly-HEMA-coated surface were compared with those in the sandwich model. Major liver-specific functions, such as albumin secretion and urea synthesis, were evaluated in both the spheroid and sandwich model. The synthesis performance in the spheroid compared to the sandwich culture increases approximately by a factor of 1.5. Disintegration of plasma membranes in both models was measured by lactate dehydrogenase (LDH) release in both models. Additionally, diazepam was used as a substrate in drug metabolism studies to characterize the differences in the biotransformation potential with metabolite profiles in both models. It showed that the diazepam metabolism activities in the spheroid model is about 10-fold lower than the sandwich model. The poly-HEMA-based hepatocyte spheroid is a promising new platform towards hepatic tissue engineering leading to in vitro hepatic tissue formation.

show abstract

Micropatterned organoid culture of rat hepatocytes and HepG2 cells

Cited by 46 publications

References 27 publications

A Strategy Using Pluripotent Stem Cell-Derived Hepatocytes for Stem Cell-Based Therapies

A Strategy Using Pluripotent Stem Cell-Derived Hepatocytes for Stem Cell-Based Therapies

Comparative Analysis of Gene Expression in Rat Liver Tissue and Monolayer- and Spheroid-Cultured Hepatocytes

Morphological and Functional Analysis of Hepatocyte Spheroids Generated on Poly-HEMA-Treated Surfaces under the Influence of Fetal Calf Serum and Nonparenchymal Cells

Contact Info

Product

Resources

About