Rapid Formation of Multicellular Spheroids of Adult Rat Hepatocytes by Rotation Culture and Their Immobilization within Calcium Alginate

Yagi, Kiyohito; Tsuda, Kozo; Serada, Masashi; Yamada, Chieko; Kondoh, A.; Miura, Yoshiharu

doi:10.1111/j.1525-1594.1993.tb00405.x

Cited by 57 publications

(7 citation statements)

References 17 publications

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“…The material chosen here, sodium alginate, is widely used for cells encapsulation both in vitro [29–31] and in vivo [32,33]. Even though unmodified alginate-based hydrogel does not support interaction with cells directly [34] it is possible to use them as a scaffold for cell immobilization that can lead to aggregation [35]. However, if needed, alginate can be modified with specific cell attachment proteins or blended with different biomaterials such as silk fibroin to improve cell adhesion [36,37].…”

Section: Discussionmentioning

confidence: 99%

Optimization of 3D bioprinting of human neuroblastoma cells using sodium alginate hydrogel

Lewicki

Bergman

Kerins

et al. 2019

Preprint

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2627 Abbreviations: FRESH, freeform reversible embedding of suspended hydrogels; POI, 28 parameter optimization index; SA, sodium alginate; 2 29 Abstract 30 There are many parameters in extrusion-based three-dimensional (3D) bioprinting of 31 different materials that require fine-tuning to obtain the optimal print resolution and cell 32 viability. To standardize this process, methods such as parameter optimization index (POI) 33 have been introduced. The POI aims at pinpointing the optimal printing speed and pressure 34 to achieve the highest accuracy keeping theoretical shear stress low. Here we applied the 35 POI to optimize the process of 3D bioprinting human neuroblastoma cell-laden 2% sodium 36 alginate (SA) hydrogel using freeform reversible embedding of suspended hydrogels (FRESH).37 Our results demonstrate a notable difference between optimal parameters for printing 2% 38 SA with and without cells in the hydrogel. We also detected a significant influence of long-39 term cell culture on the printed constructs. This observation suggests that the POI has to be 40 evaluated in the perspective of the final application. When taking these conditions into 41 consideration, we could define a set of parameters that resulted in good quality prints 42 maintaining high neuroblastoma cell viability (83% viable cells) during 7 days of cell culture 43 using 2% SA and FRESH bioprinting. These results can be further used to manufacture 44 neuroblastoma in vitro 3D culture systems to be used for cancer research. 45 3 46 2. Introduction 4748 Stem cell and tumor biology allow for the generation of small organ-like or tumor-like 49 structures to be developed in vitro, and this holds great promise for significant improvement 50 of approaches in drug discovery and precision medicine. Bioprinting has emerged as an 51 important tool for improving the conditions and control of such cell culture rationale [1,2].52 However, to achieve optimal results, many parameters of the microenvironment must be 53 taken into account. We and others have demonstrated the significance of, e.g., oxygen levels 54 [3,4], substrate stiffness [5,6], substrate roughness [7], and biomaterial properties [8] for 55 progenitor cells to respond properly to external signaling factors such as growth factors, and 56 to execute the appropriate transcriptional programs. Yet, 2-dimensional cell culture is in 57 itself a limiting factor both in stem cell and tumor biology and it has been shown that, for 58 example, certain tumor cells grown in 2D conditions are more sensitive to chemotherapeutic 59 reagents than when grown in three dimensions [9], which may explain some of the lack of 60 progress in cancer research heavily debated during recent years [10,11]. 6162 Three-dimensional (3D) bioprinting was first reported to deposit viable cells by Smith et al in 63 2004 [12]. More than a decade later, 3D bioprinting is constantly being improved in terms of 64 hardware, software, biomaterials, and applications. Standard 3D extrusion-based 65 bioprinters, despite being ...

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Section: Discussionmentioning

confidence: 99%

Optimization of 3D bioprinting of human neuroblastoma cells using sodium alginate hydrogel

Lewicki

Bergman

Kerins

et al. 2019

Preprint

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“…Also, alginate polymer is stable in culture for weeks yet can be easily depolymerized to release the cells within simply by lowering the calcium concentration. [26][27][28][29] In the past, alginate encapsulation has been used for cultures of hepatocytes and for cell lines. 30,31 Recently Chandrasekaran et al 18 showed that mouse hepatic progenitor cells cultured in alginate produce viable, long-term cultures.…”

Section: Discussionmentioning

confidence: 99%

Mature Human Hepatocytes fromEx VivoDifferentiation of Alginate-Encapsulated Hepatoblasts

Cheng

Wauthier

Reíd

2008

Tissue Engineering Part A

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Alginate gel was used to provide encapsulation to support the growth and eventually the differentiation of hepatic progenitors cells derived from human fetal livers. The encapsulated cells aggregated into spheroids within a few days in culture and continued to grow for at least 4 weeks in a serum-free medium. The hepatic progenitor cells in the spheroids undergo differentiation, as indicated by the appearance of functions of mature hepatocytes such as the detoxification of ammonia, albumin secretion, expression of the adult cytochrome P450 isozyme CYP3A4 and enzymatic activity typical of CYP2C9. Along with the expression of mature hepatic markers, these progenitor cells lost features typical of immature liver cells such as epithelial cell adhesion molecule. In addition to the acquisition of mature biochemical functions, the spheroids also developed a bile ducts, suggesting that they had differentiated into tissues resembling those in an intact liver.

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“…8), all three extracellular matrix components investigated, namely collagen type I, collagen type II, and aggrecan were identified in the beads qualitatively. Given the hydrophilicity of alginate and consequent lack of cell adhesion to the material,53–55 cells commonly form aggregates when encapsulated in alginate,55, 58 and the presence of the extracellular matrix components was generally observed qualitatively in the micrographs surrounding clusters of cells (Fig. 8).…”

Section: Discussionmentioning

confidence: 99%

Chondrogenic differentiation of neonatal human dermal fibroblasts encapsulated in alginate beads with hydrostatic compression under hypoxic conditions in the presence of bone morphogenetic protein‐2

Singh

Pierpoint

Mikos

et al. 2011

J Biomedical Materials Res

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In the present work, neonatal human dermal fibroblasts (nHDFs) were evaluated as a potential cell source for intervertebral disc repair. Chondrogenic differentiation of nHDFs was studied in the presence or absence of hydrostatic compression under normal and hypoxic conditions. In addition, the potentially synergistic effects of mechanical stimulation and bone morphogenetic protein (BMP)-2 on the chondrogenic differentiation of nHDFs were assessed. Mechanical stimulation was applied to the cells encapsulated in alginate beads using a custom-built bioreactor system for either a 1- or 3-week period at a frequency of 1 Hz for 4 h/day. In general, after 21 days of culture, high cell viability was observed for all the groups, with the exception of the groups exposed to intermittent mechanical stimulation for 3 weeks. Long-term intermittent application of mechanical stimulation under low O(2) conditions resulted in elevated collagen biosynthesis rate from day 0. Inclusion of BMP-2 for this group improved the chondrogenic differentiation of nHDFs, as indicated by elevated aggrecan gene expression and an increased collagen production rate compared to the day 0 group. Thus, the combination of hypoxia, BMP-2 supplementation, and long-term intermittent application of dynamic hydrostatic pressure was found to be a potent promoter of the chondrogenic differentiation of nHDFs.

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Rapid Formation of Multicellular Spheroids of Adult Rat Hepatocytes by Rotation Culture and Their Immobilization within Calcium Alginate

Cited by 57 publications

References 17 publications

Optimization of 3D bioprinting of human neuroblastoma cells using sodium alginate hydrogel

Optimization of 3D bioprinting of human neuroblastoma cells using sodium alginate hydrogel

Mature Human Hepatocytes fromEx VivoDifferentiation of Alginate-Encapsulated Hepatoblasts

Chondrogenic differentiation of neonatal human dermal fibroblasts encapsulated in alginate beads with hydrostatic compression under hypoxic conditions in the presence of bone morphogenetic protein‐2

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