A 3D Sphere Culture System Containing Functional Polymers for Large-Scale Human Pluripotent Stem Cell Production

Otsuji, Tomomi G.; Jiang, Bin; Yoshimura, Azumi; Tomura, Misayo; Tateyama, Daiki; Minami, Itsunari; Yoshikawa, Yusuke; Aiba, Keisuke; Heuser, John E.; Nishino, Taito; Hasegawa, Kouichi; Nakatsuji, Norio

doi:10.1016/j.stemcr.2014.04.013

Cited by 20 publications

(30 citation statements)

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“…Because it allows the 3D culture of spheroids without the requirement for stirring, FP001 is used for the 3D culture of human iPS cells. 12 Corresponding to the results of previous studies, HepaRG spheroids that formed in the medium containing FP001 demonstrated a greater level of mature hepatic characteristics than that of 2D cultured HepaRG cells. Furthermore, the comparison of gene expression patterns and albumin-expressing hepatocytes revealed that 3D culture accelerated the differentiation of HepaRG cells into hepatocytes rather than cholangiocytes in vitro and in vivo.…”

Section: Introductionsupporting

confidence: 75%

Functional polymer‐dependent 3D culture accelerates the differentiation of HepaRG cells into mature hepatocytes

Higuchi

Kawai

Kanaki

et al. 2016

Hepatology Research

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Aim:The hepatoma-derived cell line HepaRG is regarded as an in vitro model of drug metabolism because fully differentiated HepaRG cells demonstrate functional metabolic responses comparable to those of primary human hepatocytes. Recently, it was demonstrated that the 3D culture of HepaRG cells enhanced their metabolic functions and toxicological responses. We approached the mechanisms underlying these enhancement effects.Methods: We compared 2D-cultured HepaRG cells with 3D-cultured HepaRG spheroids in the gene expression patterns and the metabolic functions. In the present study, we performed 3D culture of HepaRG cells using functional polymers (FP). To reveal the in vivo differentiation ability, we transplanted the 3D-cultured HepaRG spheroids into TK-NOG mice.Results: A comparison between 2D and 3D cultures revealed that 3D-cultured HepaRG spheroids demonstrated reductions in bile duct marker expression, accelerated expression of cytochrome P450 3A4, and increases in the ratio of albuminexpressing hepatocytes. Furthermore, catalytic activities of cytochrome P450 3A4 were modified by omeprazole and rifampicin in the 3D-cultured HepaRG spheroids. Transplantation analysis revealed that 3D-cultured HepaRG spheroids formed hepatocyte-like colonies rather than cholangiocytes in vivo. Conclusion:Our results indicated that the enhancement of hepatic functions in 3D-cultured HepaRG cells was induced by selective hepatocyte differentiation and accelerated hepatocyte maturation. HepaRG spheroids reproduced the metabolic responses of human hepatocytes. Therefore, FP-dependent 3D-cultured HepaRG cells may serve as an excellent in vitro model for evaluating the hepatic metabolism and toxicity.

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Section: Introductionsupporting

confidence: 75%

Functional polymer‐dependent 3D culture accelerates the differentiation of HepaRG cells into mature hepatocytes

Higuchi

Kawai

Kanaki

et al. 2016

Hepatology Research

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“…In vitro 3D culture systems offer a solution to these limitations (3)(4)(5)(6)(7). Compared with 2D culture, a 3D arrangement allows neuronal cells to interact with many more cells (4); this similarity to the in vivo setting has been shown to lengthen viability, enhance survival, and allow formation of longer neurites and more dense networks in primary neurons in uniform matrices or aggregate culture (8,9).…”

mentioning

confidence: 99%

Layered hydrogels accelerate iPSC-derived neuronal maturation and reveal migration defects caused by MeCP2 dysfunction

Zhang

Freitas

Hao

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

142

119

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Probing a wide range of cellular phenotypes in neurodevelopmental disorders using patient-derived neural progenitor cells (NPCs) can be facilitated by 3D assays, as 2D systems cannot entirely recapitulate the arrangement of cells in the brain. Here, we developed a previously unidentified 3D migration and differentiation assay in layered hydrogels to examine how these processes are affected in neurodevelopmental disorders, such as Rett syndrome. Our soft 3D system mimics the brain environment and accelerates maturation of neurons from human induced pluripotent stem cell (iPSC)-derived NPCs, yielding electrophysiologically active neurons within just 3 wk. Using this platform, we revealed a genotype-specific effect of methyl-CpG-binding protein-2 (MeCP2) dysfunction on iPSC-derived neuronal migration and maturation (reduced neurite outgrowth and fewer synapses) in 3D layered hydrogels. Thus, this 3D system expands the range of neural phenotypes that can be studied in vitro to include those influenced by physical and mechanical stimuli or requiring specific arrangements of multiple cell types.3D hydrogels | neuronal migration and maturation | 3D RTT modeling N euronal migration and maturation is a key step in brain development. Defects in this process have been implicated in many disorders, including autism (1) and schizophrenia (2). Thoroughly understanding how neural progenitor cell (NPC) migration is affected in neurodevelopmental disorders requires a means of dissecting the process using cells with genetic alterations matching those in patients. Existing in vitro assays of migration generally involve measurement of cell movement across a scratch or gap or through a membrane toward a chemoattractant in 2D culture systems. Although widely used, such assays may not accurately reveal in vivo differences, as neuronal migration is tightly regulated by physical and chemical cues in the extracellular matrix (ECM) that NPCs encounter as they migrate.In vitro 3D culture systems offer a solution to these limitations (3-7). Compared with 2D culture, a 3D arrangement allows neuronal cells to interact with many more cells (4); this similarity to the in vivo setting has been shown to lengthen viability, enhance survival, and allow formation of longer neurites and more dense networks in primary neurons in uniform matrices or aggregate culture (8, 9). Indeed, 3D culture systems have been used to study nerve regeneration, neuronal and glial development (10-12), and amyloid-β and tau pathology (13). Thus, measuring neuronal migration through a soft 3D matrix would continue this trend toward using 3D systems to study neuronal development and pathology.We sought to develop a 3D assay to examine potential migration and neuronal maturation defects in Rett syndrome (RTT), a genetic neurodevelopmental disorder that affects 1 in 10,000 children in the United States and is caused by mutations in the X-linked methyl-CpG-binding protein-2 (MECP2) gene (14). Studies using induced pluripotent stem cells (iPSCs) from RTT patients in traditiona...

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“…PSCs may prove to be better since they have excellent proliferation and a shorter doubling time than adult stem cells. Moreover, in a 3D spheroid culture system, the PSCs may be propagated to 1 × 10 6 cells/ml in a 200‐ml plastic bag within 5 days when ∼2 × 10 8 cells in spheroids could be obtained . Kempf et al.…”

Section: Cell‐based Therapies For MImentioning

confidence: 99%

“…It is easier to ensure better quality control with PSCs than MSCs because comprehensive background information can be obtained . Moreover, PSCs can be completely manipulated with xeno‐free, well‐defined conditions in a high‐grade, GMP‐compliant facility with pipeline production for clinical use …”

Section: Cell‐based Therapies For MImentioning

confidence: 99%

Stem Cell Therapy of Myocardial Infarction: A Promising Opportunity in Bioengineering

Jiang

Shamul

et al. 2020

Advanced Therapeutics

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Myocardial infarction (MI) is a life‐threatening disease resulting from the irreversible death of cardiomyocytes (CMs). Stem cell‐based therapies have been studied for MI treatment over the last two decades with promising outcomes. Here, the past work in this field is critically reviewed to elucidate the advantages and disadvantages of treating MI using pluripotent stem cells (PSCs) including both embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), adult stem cells, and cardiac progenitor cells. Overall, PSCs (particularly iPSCs) have more advantages than the other cells for cardiac regeneration. However, the use of iPSCs is also facing critical challenges, which may be resolved by using biomaterials to engineer stem cells for reduced immunogenicity, improved immobilization/survival in the heart, and increased integration with the host cardiac tissue to eliminate arrhythmia. Biomaterials have also been applied in the derivation of CMs in vitro to increase the efficiency and maturation of cardiac differentiation. Collectively, a lot has been learned from the past failures of simply injecting intact stem cells or their derivatives in vivo for treating MI, and bioengineering stem cells with biomaterials may be a valuable strategy for advancing stem cell therapy towards its widespread application for MI treatment in the clinic.

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A 3D Sphere Culture System Containing Functional Polymers for Large-Scale Human Pluripotent Stem Cell Production

Cited by 20 publications

References 0 publications

Functional polymer‐dependent 3D culture accelerates the differentiation of HepaRG cells into mature hepatocytes

Functional polymer‐dependent 3D culture accelerates the differentiation of HepaRG cells into mature hepatocytes

Layered hydrogels accelerate iPSC-derived neuronal maturation and reveal migration defects caused by MeCP2 dysfunction

Stem Cell Therapy of Myocardial Infarction: A Promising Opportunity in Bioengineering

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