Nonadhesive Alginate Hydrogels Support Growth of Pluripotent Stem Cell-Derived Intestinal Organoids

Capeling, Meghan M.; Czerwinski, Michael; Huang, Sha; Tsai, Yu-Hwai; Wu, Angeline; Nagy, Melinda S.; Juliar, Benjamin A.; Sundaram, Nambirajan; Song, Yang; Han, Woojin M.; Takayama, Shuichi; Alsberg, Eben; Garcı́a, Andrés J.; Helmrath, Michael A.; Putnam, Andrew J.; Spence, Jason R.

doi:10.1016/j.stemcr.2018.12.001

Cited by 175 publications

(179 citation statements)

References 46 publications

Supporting

Mentioning

177

Contrasting

Order By: Relevance

“…It has been demonstrated that several niche factors allow adult and developing human and murine intestinal epithelium to be cultured ex vivo as organoids (Capeling et al, 2019; Finkbeiner et al, 2015; Fordham et al, 2013; Hill et al, 2017; Kraiczy et al, 2017; Sato et al, 2009, 2011). These factors often include WNT and RSPO ligands, BMP/TGFβ antagonists and EGF, and are based on defined growth conditions that allow expansion of intestinal epithelium in vitro (Sato et al, 2009, 2011).…”

Section: Resultsmentioning

confidence: 99%

“…The stem cell niche within a tissue is required to regulate stem cell maintenance, self-renewal and differentiation (Scadden, 2006). The niche is made up of both physical and chemical cues, including the extracellular matrix (ECM), cell-cell contacts, growth factors and other small molecules such as metabolites (Capeling et al, 2019; Cruz-Acuña et al, 2017; Gjorevski et al, 2016). Understanding the niche within various tissues has been central to understanding how tissues are maintain homeostasis, and for understanding how disease may occur (Van de Wetering et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

“…Although a wealth of information about the signaling environment within the ISC niche has informed our understanding of ISC regulation, and that this information has been leveraged to grow epithelium-only intestinal organoids (herein referred to as enteroids) in vitro , it is also clear that current in vitro systems are still not optimized to most accurately reflect the in vivo environment. Efforts have been ongoing to improve the in vitro physical environment through biomimetic ECM (Capeling et al, 2019; Cruz-Acuña et al, 2017; Gjorevski et al, 2016), and by adjusting signaling cues to more accurately reflect the in vivo niche (Fujii et al, 2018). More recently, single cell technologies have started to reveal unprecedented amounts of information about the cellular heterogeneity of human tissue and the ISC niche during health and disease (Kinchen et al, 2018; Martin et al, 2019; Smillie et al, 2019), and will undoubtedly yield substantial information about cell types and niche cues that regulate ISCs in various contexts.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

In vitroandin vivodevelopment of the human intestinal niche at single cell resolution

Czerwinski

Holloway

Tsai

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

SUMMARYThe human intestinal stem cell (ISC) niche supports ISC self-renewal and epithelial function, yet little is known about the development of the human ISC niche. We used single-cell mRNA sequencing (scRNA-seq) to interrogate the human intestine across 7-21 weeks of gestation. Using these data coupled with marker validation in situ, molecular identities and spatial locations were assigned to several cell populations that comprise the epithelial niche, and the cellular origins of many niche factors were determined. The major source of WNT and RSPONDIN ligands were ACTA2+ cells of the muscularis mucosa. EGF was predominantly expressed in the villus epithelium and the EGF-family member NEUREGULIN1 (NRG1) was expressed by subepithelial mesenchymal cells. Functional data from enteroid cultures showed that NRG1 improved cellular diversity, enhanced the stem cell gene signature, and increased enteroid forming efficiency, whereas EGF supported a secretory gene expression profile and stimulated rapid proliferation. This work highlights unappreciated complexities of intestinal EGF/ERBB signaling and identifies NRG1 as a stem cell niche factor.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

In vitroandin vivodevelopment of the human intestinal niche at single cell resolution

Czerwinski

Holloway

Tsai

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Human intestinal organoids (HIOs) have been almost exclusively characterized following growth for several weeks in culture (Capeling et al, 2019; Finkbeiner et al, 2015b; Spence et al, 2010; Tsai et al, 2017). Through these analyses, various intestinal epithelial and mesenchymal populations have been identified.…”

Section: Resultsmentioning

confidence: 99%

Differentiation of human intestinal organoids with endogenous vascular endothelial cells

Holloway

Czerwinkski

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

22Human pluripotent stem cell (hPSC)-derived intestinal organoids (HIOs) generated using directed 23 differentiation lack some cellular populations found in the native organ, including vasculature. Using 24 single cell RNA sequencing (scRNAseq), we have identified a transient population of endothelial cells 25 (ECs) present early in HIO differentiation that are lost over time in culture. Here, we have developed a 26 method to enhance co-differentiation and maintenance of ECs within HIOs (vHIOs). Given that ECs are 27 known to possess organ specific gene expression, morphology and function, we used bulk RNAseq 28 and scRNAseq to interrogate the developing human intestine, lung, and kidney in order to identify 29 organ-enriched EC-gene signatures in these organ systems. By comparing organ-specific gene 30 signatures along with markers validated by fluorescent in situ hybridization to HIO ECs, we find that 31 HIO ECs grown in vitro share the highest similarity with native intestinal ECs relative to kidney and 32 lung. Together, these data show that HIOs can co-differentiate a native EC population that are properly 33 patterned with an intestine-specific EC transcriptional signature in vitro. 34 35 36 37 38 39 40 41 42 43 44 65highly vascularized regions of immunocompromised mice (Cortez et al., 2018; Watson et al., 2014). In 66 these environments, HIOs undergo extensive vascularization by the murine host tissue, and increase in 67 complexity to resemble mature intestinal tissue (Finkbeiner et al., 2015b; Watson et al., 2014). 68However, co-culture approaches or co-differentiating HIOs with a native vasculature prior to in vivo 69 engraftment has not yet been achieved. 71Here, we performed single cell RNA sequencing (scRNAseq) at various timepoints across HIO 72 differentiation in vitro and observed a transient population of endothelial-like cells (ECs) present within 73 HIOs early during differentiation; however, these cells are not maintained during prolonged culture 74 under standard growth conditions. This suggested that early during HIO differentiation, cells within the 75 culture are capable of giving rise to EC-like cells. Based on these observations, we hypothesized that a 76 modified directed differentiation approach would allow the induction and maintenance of a more robust 77 EC population within HIOs (termed vHIO). Our findings demonstrate that modified culture conditions 78 allow a ~13-fold increase in the induction of EC-like cells within HIOs without impacting the other HIO 79 cell populations present (i.e. epithelium, mesenchyme), and support the survival of this population of 80 ECs within HIOs in culture for months. 82Since organ-specific morphology in vascular beds has long been appreciated (Aird, 2007), and 83 organ-specific transcriptional signatures and functions have been described in mouse (Ding et al., 84 2011; Kalucka et al., 2020; Lee et al., 2014; Nolan et al., 2013) and human tissues (Chi et al., 2003; 85 Marcu et al., 2018), we further sought to determine if HIO ECs were properly...

show abstract

“…Supplementation of laminin‐111 in fibrin‐based hydrogels can achieve analogous colony formation efficiency to that of the Matrigel in organoid culture. Although other hydrogels like alginate hydrogels can support the survival of intestinal organoids (Capeling et al, ), their abilities to generate well‐differentiated organoids are poorer possibly due to the lack of coupled RGD and additive laminin‐111.…”

Section: Biomaterials For Intestinal Organoid Culturementioning

confidence: 99%

Biomaterials and biosensors in intestinal organoid culture, a progress review

Huang

Jiang

Ren

et al. 2020

J Biomedical Materials Res

View full text Add to dashboard Cite

As an emerging technology, intestinal organoids are promising new tools for basic and translational research in gastroenterology. Currently, culture of intestinal organoids relies mostly on a type of tumor-derived scaffolds, namely Matrigel, which may pose tumorigenic risks to organoid implantation. Apart from the traditional detection methods, such as tissue slicing and fluorescence staining, the monitoring of intestinal organoids requires real-time biosensors that can adapt to their threedimensional dynamic growth patterns. In this review, we summarized the recent advances in developing definite hydrogel scaffolds for intestinal organoid culture and identified key parameters for scaffold design. In addition, classified by different substrate compositions like pH, electrolytes, and functional proteins, we concluded the existing live-imaging biosensors and elucidated their underlying mechanisms. We hope this review enhances the understanding of intestinal organoid culture and provides more practical approaches to investigate them. K E Y W O R D Sbiosensor, intestinal stem cell, organoid, regenerative medicine, scaffold design

show abstract

Nonadhesive Alginate Hydrogels Support Growth of Pluripotent Stem Cell-Derived Intestinal Organoids

Cited by 175 publications

References 46 publications

In vitroandin vivodevelopment of the human intestinal niche at single cell resolution

In vitroandin vivodevelopment of the human intestinal niche at single cell resolution

Differentiation of human intestinal organoids with endogenous vascular endothelial cells

Biomaterials and biosensors in intestinal organoid culture, a progress review

Contact Info

Product

Resources

About