Human iPS cell-derived fibroblast-like cells as feeder layers for iPS cell derivation and expansion

Du, Shouhui; Tay, Johan Chin-Kang; Chen, Can; Tay, Felix-Chang; Tan, Wee-Kiat; Li, Zhendong; Wang, Shu

doi:10.1016/j.jbiosc.2014.12.009

Cited by 12 publications

(5 citation statements)

References 30 publications

(34 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Importantly, we show that similar to murine GAeFib and eECs, human growth‐arrested fibroblasts and endothelial cells are also highly permissive for the specification of hiPSCs toward Pax7 + myogenic progenitors. Moreover, recent protocols allow for the generation of fibroblast‐like cells and endothelial cells from hiPSCs (Du et al , 2015; Patsch et al , 2015). Future studies addressing heterotypic suspension embryoids containing support cells derived from autologous or non‐immunogenic hiPSCs will further facilitate translational applications.…”

Section: Discussionmentioning

confidence: 99%

An engineered multicellular stem cell niche for the 3D derivation of human myogenic progenitors from iPSCs

et al. 2022

View full text Add to dashboard Cite

Fate decisions in the embryo are controlled by a plethora of microenvironmental interactions in a three-dimensional niche. To investigate whether aspects of this microenvironmental complexity can be engineered to direct myogenic human-induced pluripotent stem cell (hiPSC) differentiation, we here screened murine cell types present in the developmental or adult stem cell niche in heterotypic suspension embryoids. We identified embryonic endothelial cells and fibroblasts as highly permissive for myogenic specification of hiPSCs. After two weeks of sequential Wnt and FGF pathway induction, these three-component embryoids are enriched in Pax7positive embryonic-like myogenic progenitors that can be isolated by flow cytometry. Myogenic differentiation of hiPSCs in heterotypic embryoids relies on a specialized structural microenvironment and depends on MAPK, PI3K/AKT, and Notch signaling. After transplantation in a mouse model of Duchenne muscular dystrophy, embryonic-like myogenic progenitors repopulate the stem cell niche, reactivate after repeated injury, and, compared to adult human myoblasts, display enhanced fusion and lead to increased muscle function. Altogether, we provide a two-week protocol for efficient and scalable suspension-based 3D derivation of Pax7positive myogenic progenitors from hiPSCs.

show abstract

Section: Discussionmentioning

confidence: 99%

An engineered multicellular stem cell niche for the 3D derivation of human myogenic progenitors from iPSCs

et al. 2022

View full text Add to dashboard Cite

show abstract

“…iPS cells can be generated from various tissue sources, including fibroblasts from skin ( Alawad et al, 2016 ; Du et al, 2015 ) and hematopoietic cells ( Haase, Gohring & Martin, 2017 ; Takenaka et al, 2010 ; Kim, Manzar & Zavazava, 2013 ) or lymphocytes ( Phillips et al, 2012 ) from peripheral blood, and have been used in disease modeling. HuiPSC lines derived from peripheral blood T lymphocytes reportedly differentiate into hepatocytes more efficiently than hiPSC clones derived from adult dermal fibroblasts ( Kajiwara et al, 2012 ).…”

Section: Discussionmentioning

confidence: 99%

Generation of human liver organoids from pluripotent stem cell-derived hepatic endoderms

Kulkeaw

Tubsuwan

Tongkrajang

et al. 2020

PeerJ

View full text Add to dashboard Cite

Background The use of a personalized liver organoid derived from human-induced pluripotent stem cells (HuiPSCs) is advancing the use of in vitro disease models for the design of specific, effective therapies for individuals. Collecting patient peripheral blood cells for HuiPSC generation is preferable because it is less invasive; however, the capability of blood cell-derived HuiPSCs for hepatic differentiation and liver organoid formation remains uncertain. Moreover, the currently available methods for liver organoid formation require a multistep process of cell differentiation or a combination of hepatic endodermal, endothelial and mesenchymal cells, which is a major hurdle for the application of personalized liver organoids in high-throughput testing of drug toxicity and safety. To demonstrate the capability of blood cell-derived HuiPSCs for liver organoid formation without support from endothelial and mesenchymal cells. Methods The peripheral blood-derived HuiPSCs first differentiated into hepatic endoderm (HE) in two-dimensional (2D) culture on Matrigel-coated plates under hypoxia for 10 days. The HE was then collected and cultured in 3D culture using 50% Matrigel under ambient oxygen. The maturation of hepatocytes was further induced by adding hepatocyte growth medium containing HGF and oncostatin M on top of the 3D culture and incubating the culture for an additional 12–17 days. The function of the liver organoids was assessed using expression analysis of hepatocyte-specific gene and proteins. Albumin (ALB) synthesis, glycogen and lipid storage, and metabolism of indocyanine were evaluated. The spatial distribution of albumin was examined using immunofluorescence and confocal microscopy. Results CD34+ hematopoietic cell-derived HuiPSCs were capable of differentiating into definitive endoderm expressing SOX17 and FOXA2, hepatic endoderm expressing FOXA2, hepatoblasts expressing AFP and hepatocytes expressing ALB. On day 25 of the 2D culture, cells expressed SOX17, FOXA2, AFP and ALB, indicating the presence of cellular heterogeneity. In contrast, the hepatic endoderm spontaneously formed a spherical, hollow structure in a 3D culture of 50% Matrigel, whereas hepatoblasts and hepatocytes could not form. Microscopic observation showed a single layer of polygonal-shaped cells arranged in a 3D structure. The hepatic endoderm-derived organoid synthesis ALB at a higher level than the 2D culture but did not express definitive endoderm-specific SOX17, indicating the greater maturity of the hepatocytes in the liver organoids. Confocal microscopic images and quantitative ELISA confirmed albumin synthesis in the cytoplasm of the liver organoid and its secretion. Overall, 3D culture of the hepatic endoderm is a relatively fast, simple, and less laborious way to generate liver organoids from HuiPSCs that is more physiologically relevant than 2D culture.

show abstract

“…Therefore, lower extent of maturation was likely a limitation of our protocol. iPS cells can be generated from various tissue sources, including fibroblasts from skin [Alawad et al 2016;Du et al 2015] and hematopoietic cells [Haase et al 2017;Takenaka et al 2010;Kim et al 2013] or lymphocytes (Phillips et al 2012) from peripheral blood, and have been used in disease modeling. HuiPSC lines derived from peripheral blood T lymphocytes reportedly differentiate into hepatocytes more efficiently than hiPSC clones derived from adult dermal fibroblasts (Kajiwara et al 2012).…”

Section: ) Asmentioning

confidence: 99%

Peer Review #1 of "Generation of human liver organoids from pluripotent stem cell-derived hepatic endoderms (v0.1)"

2020

View full text Add to dashboard Cite

Background. The use of a personalized liver organoid derived from human-induced pluripotent stem cells (HuiPSCs) is advancing the use of in vitro disease models for the design of specific, effective therapies for individuals. Collecting patient peripheral blood cells for HuiPSC generation is preferable because it is less invasive; however, the capability of blood cell-derived HuiPSCs for hepatic differentiation and liver organoid formation remains uncertain. Moreover, the currently available methods for liver organoid formation require a multistep process of cell differentiation or a combination of hepatic endodermal, endothelial and mesenchymal cells, which is a major hurdle for the application of personalized liver organoids in high-throughput testing of drug toxicity and safety. To demonstrate the capability of blood cell-derived HuiPSCs for liver organoid formation without support from endothelial and mesenchymal cells. Methods. The peripheral blood-derived HuiPSCs first differentiated into hepatic endoderm (HE) in twodimensional (2D) culture on Matrigel-coated plates under hypoxia for 10 days. The HE was then collected and cultured in 3D culture using 50% Matrigel under ambient oxygen. The maturation of hepatocytes was further induced by adding hepatocyte growth medium containing HGF and oncostatin M on top of the 3D culture and incubating the culture for an additional 12-17 days. The function of the liver organoids was assessed using expression analysis of hepatocyte-specific gene and proteins. Albumin (ALB) synthesis, glycogen and lipid storage, and metabolism of indocyanine were evaluated. The spatial distribution of albumin was examined using immunofluorescence and confocal microscopy. Results. CD34+ hematopoietic cell-derived HuiPSCs were capable of differentiating into definitive endoderm expressing SOX17 and FOXA2, hepatic endoderm expressing FOXA2, hepatoblasts expressing AFP and hepatocytes expressing ALB. On day 25 of the 2D culture, cells expressed SOX17, FOXA2, AFP and ALB, indicating the presence of cellular heterogeneity. In contrast, the hepatic endoderm spontaneously formed a spherical, hollow structure in a 3D culture of 50% Matrigel, whereas hepatoblasts and hepatocytes could not form. Microscopic observation showed a single layer of polygonal-shaped cells arranged in a 3D structure. The hepatic endoderm-derived organoid synthesis ALB at a higher level than the 2D culture but did not express definitive endoderm-specific SOX17, indicating the greater maturity of the hepatocytes in the liver organoids. Confocal microscopic images and quantitative ELISA confirmed albumin synthesis in the cytoplasm of the liver organoid and its secretion. Overall, 3D culture of the hepatic endoderm is a relatively fast, simple, and less laborious way to generate liver organoids from HuiPSCs that is more physiologically relevant than 2D culture.

show abstract

Human iPS cell-derived fibroblast-like cells as feeder layers for iPS cell derivation and expansion

Cited by 12 publications

References 30 publications

An engineered multicellular stem cell niche for the 3D derivation of human myogenic progenitors from iPSCs

An engineered multicellular stem cell niche for the 3D derivation of human myogenic progenitors from iPSCs

Generation of human liver organoids from pluripotent stem cell-derived hepatic endoderms

Peer Review #1 of "Generation of human liver organoids from pluripotent stem cell-derived hepatic endoderms (v0.1)"

Contact Info

Product

Resources

About