Collagenase treatment enhances proteomic coverage of low-abundance proteins in decellularized matrix bioscaffolds

Kuljanin, Miljan; Brown, Cody; Raleigh, Matthew J.; Lajoie, Gilles; Flynn, Lauren E.

doi:10.1016/j.biomaterials.2017.08.012

Cited by 40 publications

(32 citation statements)

References 74 publications

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“…Many studies assessing ECM-derived biomaterials lack appropriate tissue-type controls that are structurally and biomechanically similar, to be able to interpret whether there are tissuespecific compositional effects on the cells. [41] Hence, commercially sourced bovine tendon collagen served as a tissue-type control based on our previous studies characterizing the protein composition of this specific source, [42] as well as our previous success in applying -amylase-digested COL to generate compositionally distinct porous foams relative to DAT. [32] As expected, digestion with -amylase better preserved the fibrillar structure of the ECM sources and retained a range of higher molecular weight proteins.…”

Section: Discussionmentioning

confidence: 99%

Culture on Tissue‐Specific Coatings Derived from α‐Amylase‐Digested Decellularized Adipose Tissue Enhances the Proliferation and Adipogenic Differentiation of Human Adipose‐Derived Stromal Cells

Shridhar

Lam

Sun

et al. 2019

Biotechnology Journal

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While extracellular matrix (ECM)‐derived coatings have the potential to direct the response of cell populations in culture, there is a need to investigate the effects of ECM sourcing and processing on substrate bioactivity. To develop improved cell culture models for studying adipogenesis, the current study examines the proliferation and adipogenic differentiation of human adipose‐derived stem/stromal cells (ASCs) on a range of ECM‐derived coatings. Human decellularized adipose tissue (DAT) and commercially available bovine tendon collagen (COL) are digested with α‐amylase or pepsin to prepare the coatings. Physical characterization demonstrates that α‐amylase digestion generates softer, thicker, and more stable coatings, with a fibrous tissue‐like ultrastructure that is lost in the pepsin‐digested thin films. ASCs cultured on the α‐amylase‐digested ECM have a more spindle‐shaped morphology, and proliferation is significantly enhanced on the α‐amylase‐digested DAT coatings. Further, the α‐amylase‐digested DAT provides a more pro‐adipogenic microenvironment, based on higher levels of adipogenic gene expression, glycerol‐3‐phosphate dehydrogenase (GPDH) enzyme activity, and perilipin staining. Overall, this study supports α‐amylase digestion as a new approach for generating bioactive ECM‐derived coatings, and demonstrates tissue‐specific bioactivity using adipose‐derived ECM to enhance ASC proliferation and adipogenic differentiation.

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Section: Discussionmentioning

confidence: 99%

Culture on Tissue‐Specific Coatings Derived from α‐Amylase‐Digested Decellularized Adipose Tissue Enhances the Proliferation and Adipogenic Differentiation of Human Adipose‐Derived Stromal Cells

Shridhar

Lam

Sun

et al. 2019

Biotechnology Journal

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“…Components of the adipose extracellular matrix, such as glycosaminoglycans and proteoglycans possess the ability to bind soluble growth factors and cytokines, which allow the ECM to serve as a reservoir of these biologically active factors [47,48]. A recent mass spectrometric analysis of adipose ECM revealed that it contains a variety of matricellular proteins and adhesion glycoproteins known to be key modulators of cell functions, as well as a range of growth factors [49]. Therefore, from our study, it is reasonable to speculate that the mechanical emulsification of adipose tissue led to a rapid rise in bioactive factors that were originally stored in cells and ECM of adipose tissue, and these factors were then collected as ALE after centrifugation of the emulsified adipose tissue.…”

Section: Discussionmentioning

confidence: 99%

Human adipose liquid extract induces angiogenesis and adipogenesis: a novel cell-free therapeutic agent

Xia

Chen

et al. 2019

Stem Cell Res Ther

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Background Taking advantage of cellular paracrine mechanisms, the secretome of adipose-derived stem cells (ADSCs) and adipose tissue has been demonstrated to induce tissue repair and regeneration in various ischemic and impaired conditions. However, these cell-based therapies have been hindered by issues, such as inherent safety and cost-efficiency for clinical applications. In this study, we prepared a liquid cell-free extract from human adipose tissue [adipose liquid extract (ALE)] and evaluated its potential therapeutic efficacy. Methods ALE was prepared from human subcutaneous adipose tissue using a rapid and physical approach, and the protein components in ALE were identified using mass spectrometry analysis. In vivo, the therapeutic effect of this agent was investigated on wound healing in C57BL/6 mice, and wound healing rate, vessel density, and neo-adipocyte formation in wounded skins were measured at days 3, 7, 11, and 14. In vitro, the effect of ALE on the viability of human ADSCs, tube formation of human umbilical vein endothelial cells (HUVECs), and adipogenic differentiation of ADSCs were tested. Results The results demonstrated that ALE contained a variety of growth factors and did not affect cell viability. ALE-treated wounds exhibited accelerated wound healing with increased vessel density and formation of neo-adipocytes compared to that of control wounds. Moreover, when added as a cell culture supplement, ALE effectively induced tube formation of HUVECs and lipid accumulation in ADSCs. ALE-treated ADSCs also exhibited elevated levels of adipogenic gene expression. Conclusions ALE is a novel growth-rich therapeutic agent that is cell-free and easy to produce. Besides, it is also able to induce angiogenesis and adipogenesis both in vitro and in vivo, thus indicating that it could be used for wound repair and soft tissue regeneration.

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“…In addition, the depletion of highly abundant ECM elements such as collagens, through treatment with collagenases, can enhance the proteomic coverage of low‐abundance proteins in decellularized tissues (Kuljanin et al . ).…”

Section: Proteomic Cataloguing Of the Ecmmentioning

confidence: 97%

Charting the unexplored extracellular matrix in cancer

Filipe

Chitty

Cox

2018

Int J Experimental Path

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The extracellular matrix (ECM) is present in all solid tissues and considered a master regulator of cell behaviour and phenotype. The importance of maintaining the correct biochemical and biophysical properties of the ECM, and the subsequent regulation of cell and tissue homeostasis, is illustrated by the simple fact that the ECM is highly dysregulated in many different types of disease, especially cancer. The loss of tissue ECM homeostasis and integrity is seen as one of the hallmarks of cancer and typically defines transitional events in progression and metastasis. The vast majority of cancer studies place an emphasis on exploring the behaviour and intrinsic signalling pathways of tumour cells. Their goal was to identify ways to target intracellular pathways regulating cancer. Cancer progression and metastasis are powerfully influenced by the ECM and thus present a vast, unexplored repository of anticancer targets that we are only just beginning to tap into. Deconstructing the complexity of the tumour ECM landscape and identifying the interactions between the many cell types, soluble factors and extracellular-matrix proteins have proved challenging. Here, we discuss some of the emerging tools and platforms being used to catalogue and chart the ECM in cancer.

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Collagenase treatment enhances proteomic coverage of low-abundance proteins in decellularized matrix bioscaffolds

Cited by 40 publications

References 74 publications

Culture on Tissue‐Specific Coatings Derived from α‐Amylase‐Digested Decellularized Adipose Tissue Enhances the Proliferation and Adipogenic Differentiation of Human Adipose‐Derived Stromal Cells

Culture on Tissue‐Specific Coatings Derived from α‐Amylase‐Digested Decellularized Adipose Tissue Enhances the Proliferation and Adipogenic Differentiation of Human Adipose‐Derived Stromal Cells

Human adipose liquid extract induces angiogenesis and adipogenesis: a novel cell-free therapeutic agent

Charting the unexplored extracellular matrix in cancer

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