Natural extracellular matrix scaffolds recycled from human salivary digests: a morphometric study

Lilliu, Marzia; Seo, YJ; Isola, Michela; Charbonneau, AM; Zeitouni, Anthony; El-Hakim, Michel; Tran, SD

doi:10.1111/odi.12444

Cited by 18 publications

(20 citation statements)

References 24 publications

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“…16,24 Synthetic agarose gel–replicated organotypic scaffolds or natural ECM scaffolds were also introduced for assembly of salivary-gland microtissue. 25,26…”

Section: Discussionmentioning

confidence: 99%

Organotypic 3D Culture in Nanoscaffold Microwells Supports Salivary Gland Stem-Cell-Based Organization

Shin

Hong

Koh

et al. 2018

ACS Biomater. Sci. Eng.

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The self-organizing properties of stem cells have been exploited to generate organoids, organ-specific, cell-containing, three-dimensional (3D) structures. The present study aimed to introduce a novel bioengineering technique for driving the effective organization of adult tissue stem cells via niche-independent 3D microwell culture. Microwells were fabricated by photopatterning poly(ethylene glycol) hydrogel in the presence of an electrospun polycaprolactone nanofibrous scaffold. Human single clonal salivary gland stem cells (SGSCs) were cultured in nanofibrous microwells through two simple steps, priming and differentiation. Before the induction of 3D organization, single clonal SGSCs were preconditioned to aggregate to form 3D spheroids in different matrices, such as Matrigel, floating dish, and microwells. Expression of salivary stem cell markers and pluripotency markers was greater in 3D spheroid cultures than in 2D plastic culture. Lobular structures were organized by changing media, and those in microwells exhibited higher salivary acinar, ductal, and tight junction marker levels and decreased stem-cell marker levels relative to other 3D cultures. Furthermore, higher α-amylase secretion and intracellular calcium levels were observed in the presence of adrenergic or cholinergic agonists, respectively, along with an increased intracellular pH, suggesting more functional salivary organoid formation. These microwell-driven organoids also engrafted successfully into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. Our results showed that microwell-cultured SGSCs organize into salivary structures and that this biomimetic 3D culture technique can promote effective generation of niche-independent single stem-cell-based 3D organoids.

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“…16,24 Synthetic agarose gel–replicated organotypic scaffolds or natural ECM scaffolds were also introduced for assembly of salivary-gland microtissue. 25,26…”

Section: Discussionmentioning

confidence: 99%

Organotypic 3D Culture in Nanoscaffold Microwells Supports Salivary Gland Stem-Cell-Based Organization

Shin

Hong

Koh

et al. 2018

ACS Biomater. Sci. Eng.

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“…To date, however, the expansion of the enzymatically obtained primary acinar cells to a clinically useful mass and the maintenance of an acinar‐specific maturation state have been the two biggest technological hurdles to overcome in vitro. Thus, research efforts in the past decade have largely focused on identifying tissue‐resident stem cells that can self‐renew (Lombaert et al, ) and/or encapsulating cells in various matrix‐derived scaffolds, in which salivary epithelial cells spontaneously organize into spheroids with acinar‐like properties (Cantara et al, ; Lilliu et al, ; Maria, Liu, El‐Hakim, Zeitouni, & Tran, ; Maria, Zeitouni, Gologan, & Tran, ; Pradhan‐Bhatt et al, ; Soscia et al, ; Srinivasan et al, ).…”

Section: Introductionmentioning

confidence: 99%

“…Thus, research efforts in the past decade have largely focused on identifying tissue-resident stem cells that can self-renew (Lombaert et al, 2016) and/or encapsulating cells in various matrix-derived scaffolds, in which salivary epithelial cells spontaneously organize into spheroids with acinar-like properties (Cantara et al, 2012;Lilliu et al, 2016;Maria, Liu, El-Hakim, Zeitouni, & Tran, 2016;Maria, Zeitouni, Gologan, & Tran, 2011;Pradhan-Bhatt et al, 2013;Soscia et al, 2013;Srinivasan et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Cell culture of differentiated human salivary epithelial cells in a serum‐free and scalable suspension system: The salivary functional units model

Seo

Lilliu

Elghanam

et al. 2019

J Tissue Eng Regen Med

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Saliva aids in digestion, lubrication, and protection of the oral cavity against dental caries and oropharyngeal infections. Reduced salivary secretion, below an adequate level to sustain normal oral functions, is unfortunately experienced by head and neck cancer patients treated with radiotherapy and by patients with Sjögren's syndrome. No disease‐modifying therapies exist to date to address salivary gland hypofunction (xerostomia, dry mouth) because pharmacotherapies are limited by the need for residual secretory acinar cells, which are lost at the time of diagnosis, whereas novel platforms such as cell therapies are yet immature for clinical applications. Autologous salivary gland primary cells have clinical utility as personalized cell therapies, if they could be cultured to a therapeutically useful mass while maintaining their in vivo phenotype. Here, we devised a serum‐free scalable suspension culture system that grows partially digested human salivary tissue filtrates composing of acinar and ductal cells attached to their native extracellular matrix components while retaining their 3D in vivo spatial organization; we have coined these salivary spheroids as salivary functional units (SFU). The proposed SFU culture system was sub‐optimal, but we have found that the cells could still survive and grow into larger salivary spheroids through cell proliferation and aggregation for 5 to 10 days within the oxygen diffusion rates in vitro. In summary, by using a less disruptive cell isolation procedure as the starting point for primary cell culture of human salivary epithelial cells, we demonstrated that aggregates of cells remained proliferative and continued to express acinar and ductal cell‐specific markers.

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“…We hypothesized that the 30% cell death at day 0 (Figure ) was mainly caused by the mechanical damage during the preparation and sectioning of the slices. When compared with mechanically and enzymatically digested human and monkey salivary glands from our previous work with 2D polarized cell culture, it was not uncommon to observe that 50% of cells did not attach following plating (Tran et al , , ; Lilliu et al , ). Thus, the slice method was not harsher to cells than the conventional mechanical/enzymatic method.…”

Section: Discussionmentioning

confidence: 99%

Three‐dimensional organotypic culture of human salivary glands: the slice culture model

Fang

Liu

et al. 2016

Oral Diseases

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Natural extracellular matrix scaffolds recycled from human salivary digests: a morphometric study

Abstract: nECMsc retained native ECM proteins and maintained their distribution. Seeded cells remained viable on nECMsc.

Cited by 18 publications

References 24 publications

Organotypic 3D Culture in Nanoscaffold Microwells Supports Salivary Gland Stem-Cell-Based Organization

Organotypic 3D Culture in Nanoscaffold Microwells Supports Salivary Gland Stem-Cell-Based Organization

Cell culture of differentiated human salivary epithelial cells in a serum‐free and scalable suspension system: The salivary functional units model

Three‐dimensional organotypic culture of human salivary glands: the slice culture model

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