Perfusion culture enhanced human endometrial stromal cell growth in alginate‐multivalent integrin α5β1 ligand scaffolds

Li, Zhaohui; Kreiner, M.; Edrada-Ebel, RuAngelie; Cui, Zhanfeng; Walle, Christopher F. van der; Mardon, Helen J.

doi:10.1002/jbm.a.33177

Cited by 11 publications

(8 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…60,61 One explanation for the failure of cells to elongate and migrate in the 70% crosslink condition is that the cells must cleave a greater number of crosslinks, and are kinetically trapped, but an additional plausible explanation is that the dense crosslinks also obscure access to the adhesion ligands, and may prevent the integrin clustering required to gain sufficient signaling and traction for motility. 62–64 …”

Section: Resultsmentioning

confidence: 99%

Local remodeling of synthetic extracellular matrix microenvironments by co-cultured endometrial epithelial and stromal cells enables long-term dynamic physiological function

Cook

Hill

Guo

et al. 2017

Integrative Biology

101

View full text Add to dashboard Cite

Mucosal barrier tissues, comprising a layer of tightly-bonded epithelial cells in intimate molecular communication with an underlying matrix-rich stroma containing fibroblasts and immune cells, are prominent targets for drugs against infection, chronic inflammation, and other disease processes. Although human in vitro models of such barriers are needed for mechanistic studies and drug development, differences in extracellular matrix (ECM) needs of epithelial and stromal cells hinder efforts to create such models. Here, using the endometrium as an example mucosal barrier, we describe a synthetic, modular ECM hydrogel suitable for 3D functional co-culture, featuring components that can be remodeled by cells and that respond dynamically to sequester local cell-secreted ECM characteristic of each cell type. The synthetic hydrogel combines peptides with off-the-shelf reagents and is thus accessible to cell biology labs. Specifically, we first identified a single peptide as suitable for initial attachment of both endometrial epithelial and stromal cells using a 2D semi-empirical screen. Then, using a co-culture system of epithelial cells cultured on top of gel-encapsulated stromal cells, we show that inclusion of ECM-binding peptides in the hydrogel, along with the integrin-binding peptide, leads to enhanced accumulation of basement membrane beneath the epithelial layer and more fibrillar collagen matrix assembly by stromal cells over two weeks in culture. Importantly, endometrial co-cultures composed of either cell lines or primary cells displayed hormone-mediated differentiation as assessed by morphological changes and secretory protein production. A multiplex analysis of apical cytokine and growth factor secretion comparing cell lines and primary cells revealed strikingly different patterns, underscoring the importance of using primary cell models in analysis of cell-cell communication networks. In summary, we define a “one-size-fits-all” synthetic ECM that enables long-term, physiologically responsive co-cultures of epithelial and stromal cells in a mucosal barrier format.

show abstract

Section: Resultsmentioning

confidence: 99%

Local remodeling of synthetic extracellular matrix microenvironments by co-cultured endometrial epithelial and stromal cells enables long-term dynamic physiological function

Cook

Hill

Guo

et al. 2017

Integrative Biology

101

View full text Add to dashboard Cite

show abstract

“…Our data are consistent with a recent report on enhanced growth of human endometrial stromal cells encapsulated in alginate microbeads functionalized with multimeric fibronectin. 37 As for all MSC, establishment of optimal ex vivo growth conditions for eMSC is an important prerequisite for their potential use in cell-based therapies, since low numbers retrieved from endometrium necessitates substantial ex vivo expansion. We have previously identified four growth factors, FGF2, EGF, TGFa, and PDGF-BB, that individually supported CFU activity of freshly isolated human endometrial stromal cells in serum-free media.…”

Section: Discussionmentioning

confidence: 99%

Optimization and Scale-up Culture of Human Endometrial Multipotent Mesenchymal Stromal Cells: Potential for Clinical Application

Rajaraman

White

Tan

et al. 2013

Tissue Engineering Part C: Methods

View full text Add to dashboard Cite

We have previously identified and purified multipotent mesenchymal stromal cell (MSC)-like cells in the highly regenerative endometrial lining of the human uterus (eMSC) as CD140b⁺CD146⁺ cells. Due to ease of accessibility with minimal morbidity via biopsy, we are proposing to use eMSC in cell-based therapies; however, culture conditions compliant with Good Manufacturing Practice have not been established for eMSC. The aim of this study was to optimize serum-free and xeno-free culture conditions for expansion of eMSC for potential clinical use. Real-time cell assessment (Xcelligence) and MTS viability assays were used to measure attachment and proliferation of freshly isolated, flow cytometry-sorted CD140b⁺CD146⁺ eMSC cultured in several commercially available and in-house serum-free and xeno-free media in combination with five attachment matrices (fibronectin, collagen, gelatin, laminin, and Cell Start-XF®). Comparisons were made with a standard serum-containing medium, DMEM/F-12/10% fetal bovine serum. Under all conditions examined, eMSC attachment and proliferation was greatest using a fibronectin matrix, with Lonza TP-SF® and our in-house DMEM/SF/FGF2/EGF serum-free xeno-product-containing medium similar to serum-containing medium. Hypoxia increased eMSC proliferation in the DMEM/SF/FGF2/EGF serum-free medium. Culture of eMSC for 7 days on a fibronectin matrix in DMEM/SF/FGF2/EGF serum-free media in 5% O₂ maintained greater numbers of undifferentiated eMSC expressing CD140b, CD146, and W5C5 compared to culture under similar conditions in Lonza TP-SF medium. However, the percentage of cells expressing typical MSC phenotypic markers, CD29, CD44, CD73, and CD105, were similar for both media. EMSC showed greater expansion in 2D compared to 3D culture on fibronectin-coated microbeads using the optimized DMEM/SF/FGF2/EGF medium in 5% O₂. In the optimized 2D culture conditions, eMSC retained CFU activity, multipotency, and MSC surface phenotype, representing the first steps in their preparation for potential clinical use.

show abstract

“…better gas exchange, higher mass transfer, more physiological local solute concentrations and shear stress [35,36]. Among the dynamic culture system available on the market, TissueFlex ® , a multichannel single direction perfusion culture system, is particularly suitable for long-term culture of small-scale 3D hydrogel-based cell/tissue models [16,21]. In this study, we demonstrate that 3D encapsulation of islet is essential but perfusion culture is the key to maintain in vitro islet viability and function in culture.…”

Section: Discussionmentioning

confidence: 99%

“…Culture conditions have a profound effect on the cell viability and function in 3D scaffolds [15,16]. For instance, perfusion culture of hepatocytes retained liver metabolic functions comparable to those in vivo [17].…”

Section: Introductionmentioning

confidence: 99%

Development of In Vitro 3D TissueFlex® Islet Model for Diabetic Drug Efficacy Testing

Sun

Zhang

et al. 2013

PLoS ONE

Self Cite

View full text Add to dashboard Cite

Increasing individuals diagnosed with type II diabetes pose a strong demand for the development of more effective anti-diabetic drugs. However, expensive, ethically controversial animal-based screening for anti-diabetic compounds is not always predictive of the human response. The use of in vitro cell-based models in research presents obviously ethical and cost advantages over in vivo models. This study was to develop an in vitro three-dimensional (3D) perfused culture model of islets (Islet TF) for maintaining viability and functionality longer for diabetic drug efficacy tests. Briefly fresh isolated rat islets were encapsulated in ultrapure alginate and the encapsulated islets were cultured in TissueFlex®, a multiple, parallel perfused microbioreactor system for 7 days. The encapsulated islets cultured statically in cell culture plates (3D static) and islets cultured in suspension (2D) were used as the comparisons. In this study we demonstrate for the first time that Islet TF model can maintain the in vitro islet viability, and more importantly, the elevated functionality in terms of insulin release and dynamic responses over a 7-day culture period. The Islet TF displays a high sensitivity in responding to drugs and drug dosages over conventional 2D and 3D static models. Actual drug administration in clinics could be simulated using the developed Islet TF model, and the patterns of insulin release response to the tested drugs were in agreement with the data obtained in vivo. Islet TF could be a more predictive in vitro model for routine short- and long-term anti-diabetic drug efficacy testing.

show abstract

Perfusion culture enhanced human endometrial stromal cell growth in alginate‐multivalent integrin α5β1 ligand scaffolds

Cited by 11 publications

References 50 publications

Local remodeling of synthetic extracellular matrix microenvironments by co-cultured endometrial epithelial and stromal cells enables long-term dynamic physiological function

Local remodeling of synthetic extracellular matrix microenvironments by co-cultured endometrial epithelial and stromal cells enables long-term dynamic physiological function

Optimization and Scale-up Culture of Human Endometrial Multipotent Mesenchymal Stromal Cells: Potential for Clinical Application

Development of In Vitro 3D TissueFlex® Islet Model for Diabetic Drug Efficacy Testing

Contact Info

Product

Resources

About