Decellularized Extracellular Matrix: characterization, fabrication and

366 Pages·2019·12.648 MB·English

Decellularized Extracellular Matrix
Characterization, Fabrication and Applications
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Biomaterials Science Series
Editor-­in-c­hief:
Julian Jones, Imperial­College­London,­UK
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59 Cole DeForest, University­of­Washington,­USA
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80 Changyou Gao,­Zhejiang­University,­China
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3 1: Stimuli- responsive Drug Delivery Systems
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0. 2: Biodegradable Thermogels
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c.or 5: Antimicrobial Materials for Biomedical Applications
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Decellularized Extracellular
Matrix
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FP Characterization, Fabrication and
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1 Edited by
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org | Tetsuji Yamaoka
sc. National­Cerebral­and­Cardiovascular­Center­Research­Institute,­Japan
bs.r Email: [email protected]
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http and
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19 Takashi Hoshiba
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er Tokyo­Metropolitan­Industrial­Technology­Research­Institute,­Japan
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5 Preface
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oi: Recent progress in science and engineering, particularly in biology, pharma-
g | d cology, and materials science, has opened a new era in medicine and bio-
or engineering. In particular, expanding knowledge of the in vivo extracellular
c.
s.rs microenvironment has led to major advancements in the development of
ub pharmacological treatments for tissue and organ dysfunctions through the
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s:// use of various bioactive molecules. This knowledge has helped in the genera-
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htt tion of new cell- culture techniques that have led to innovations in cell-b ased
on therapies and bioengineering, like regenerative medicine and tissue engi-
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1 neering. For instance, cells isolated from tissues or organs can be expanded
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er in vitro via these advanced cell culture techniques and can be subsequently
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m utilized for cell transplantation therapy. In pharmaceutical science, these
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ec cultured cells have been implemented in lieu of animal models to evaluate
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5 drugs and have proven to be a suitable replacement from the perspective
n 0 of animal welfare. Various stem cells, including embryonic stem (ES) cells
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ed and induced pluripotent stem (iPS) cells, have accelerated the development
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bli of these new therapies, medicines, and bioengineering. However, in order
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P to put these cells to practical use, their cellular functions must be precisely
regulated by applying the valuable data acquired from research on the extra-
cellular microenvironment.
The extracellular microenvironment consists of different types of extra-
cellular matrices (ECMs) and various soluble factors, such as growth factors
and hormones, whose biological functions have been investigated in detail.
These factors are routinely employed as cell culture supplements for regu-
lating cell differentiation and proliferation, and some are used clinically for
wound healing, ischemia treatment, and for other therapeutic purposes. The
ECM has recently attracted a great deal of attention as biological scaffold-
ing in regenerative medicine as it supports cell survival, growth, migration,

Biomaterials Science Series No. 6
Decellularized Extracellular Matrix: Characterization, Fabrication and Applications
Edited by Tetsuji Yamaoka and Takashi Hoshiba
© The Royal Society of Chemistry 2020
Published by the Royal Society of Chemistry, www.rsc.org
v
View Online
vi Preface
morphology, and differentiation, thereby aiding tissue and organ regenera-
tion. Ideally, native ECM would be excellent scaffolds in both regenerative
medicine and cell culture. However, it is extremely difficult to reconstitute
native ECM in vitro due to its compositional, structural, and mechanical
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00 complexities. For this reason, decellularization has been widely used to
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8-F obtain biomaterials that are similar to native ECM; these biomaterials have
99 been referred to as decellularized extracellular matrix (dECM). These ECMs
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01 retain the native microstructure of tissues and organs, thereby easily allow-
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78 ing for their fabrication into anatomically-r elevant morphologies and the
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78 robust induction of tissue- specific functions. dECM, therefore provides a fit-
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1 broad fields, including regenerative medicine, tissue engineering, biological
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g | d limited due to improper preparation methods that lead to compositional,
or structural, and mechanical damage, and loss of biological activity. There are
c.
s many important points in dECM preparation, these include decellularization
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htt Fabrication, and Applications,” the recent advances in the characterization,
n fabrication, and preparation of dECM are presented. Applications of dECM
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19 in tissue engineering, regenerative medicine, clinical use, and biological
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er 2 research have been introduced by the world's leading researchers in these
mb fields. In Part I of the book, a general introduction regarding the basic knowl-
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ec edge of ECM has been presented. In Parts II, III, and IV, the preparation,
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5 characterization, and fabrication of dECM are overviewed by authors from
n 0 eight research groups, with Part II describing the classification of the dif-
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ed ferent types of dECM on the basis of their specific sources. Parts III and IV
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blis describe the characterization and fabrication of dECM, respectively. In Parts
Pu V and VI, recent examples of dECM are introduced by authors from eight
research groups, with Part V mainly focusing on tissue- and organ- derived
dECMs, and Part VI focusing on cell culture- derived dECMs.
We propose that this book will provide valuable information to research-
ers in academia and industry, as well as to postgraduate and undergraduate
students, who have an interest in dECM development. We trust that the con-
tents within this book will help to resolve the problems faced in regenera-
tive medicine, tissue engineering, pharmacology, basic biological research,
and in clinical settings. We would like to express our gratitude to all authors,
the Royal Society of Chemistry publishing team, and Yukiko Uchida of the
National Cerebral and Cardiovascular Center Research Institute for their
contributions to this book. Finally, all contributors hope that this book will
serve as a valuable resource for the scientific community.
Tetsuji Yamaoka
National Cerebral and Cardiovascular Center Research Institute, Japan
Takashi Hoshiba
Tokyo Metropolitan Industrial Technology Research Institute, Japan
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oi:1 Part I: General Introduction
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c. Chapter 1 Extracellular Matrix Scaffolds for Tissue Engineering and
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ber 1.2.2 Structures 5
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d o 1.3.2 Matrigel® (EHS Gel) 9
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ubli 1.4 Conclusion 11
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References 12
Part II: Preparation of dECM
Chapter 2 Preparation Methods for Tissue/Organ- derived
dECMs – Effects on Cell Removal and ECM Changes 17
Tetsuji Yamaoka
2.1 Introduction 17
2.2 Decellularization Methods 18

Biomaterials Science Series No. 6
Decellularized Extracellular Matrix: Characterization, Fabrication and Applications
Edited by Tetsuji Yamaoka and Takashi Hoshiba
© The Royal Society of Chemistry 2020
Published by the Royal Society of Chemistry, www.rsc.org
vii
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viii Contents
2.2.1 Physical Methods 18
2.2.2 Chemical Methods 19
2.2.3 Enzymatic Methods 20
2.3 Recellularization 21
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99 Acknowledgements 23
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n 3.2.3 Incorporation Into Assembled ECM by
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mb 3.3 Points Regarding Preparing Cultured
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n 0 3.3.2 Culture Medium and Co- culture 36
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Pu 3.3.5 Modification of Cell- derived dECM 39
3.4 Characterization of Cultured Cell- derived dECM 39
3.4.1 Confirmation of Decellularization 39
3.4.2 Identification of ECM Components 40
3.4.3 Observation of ECM Structure 41
3.5 Applications 41
3.5.1 Trials for Applications in Regenerative
Medicine and Pharmacology 41
3.5.2 Basic Biological Research 42
3.5.3 Clinical Applications 43
3.6 Perspectives 43
3.6.1 Preparation Methods 43
3.6.2 Mechanisms 44
3.6.3 Standardization for Clinical Use 44
3.7 Conclusion 45
Acknowledgements 45
References 45
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Contents ix
Chapter 4 Bared Basement Membrane Substrata: Design, Cellular
Assembly, Decellularization and Application to Tissue
Regeneration and Stem Cell Differentiation 51
Katsumi Mochitate, Reiko Nagano and Yukiko
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mb 4.5 Promotion of Basement Membrane Formation by
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Pu Assembly 59
4.5.2 Recruitment of BM Assembly
Receptor onto the Basal Surface 59
4.5.3 Assembly of a Continuous BM Architecture
on “fib” Substrata Coated with PV–GlcNAc
and –CA/Lam 63
4.5.4 Assembly of a Continuous BM Architecture
by rLN- 10 Cells on the “fib” Substratum
Coated with MAST- oligoGlcNAc: Preparation
of rLN10_sBM 67
4.6 Cellular Recognition of de novo Synthesized
Basement Membrane (sBM): Anchoring Filaments 68
4.7 Construction of Epithelial Tissue Equivalents
on sBM Substrata 69
4.7.1 Terminal Differentiation of Tracheal Basal
Cells to Ciliated Phenotype on T2- fib- MG_sBM
Substratum 70
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4.7.2 Differentiation of Embryonic Stem Cells to
Hepatocytes and Pancreatic β Cells on
rLN10_sBM 70
4.7.3 Primary Hepatocyte Culture on
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ub Pressurization and Cultured Epidermal Autograft 79
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on 5.1 Introduction 79
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of GCMN Using HHP and Grafting of CEA 85
5.4 Exploring the Degeneration of Human
Skin GCMN After HHP 86
5.5 Regression of Melanin Pigments After
Transplantation of Inactivated Nevus Tissue 88
5.6 Clinical Trial 89
5.7 Discussion 90
5.8 Conclusion 91
Acknowledgements 91
References 91
Chapter 6 Immune Responses to Decellularized Matrices 95
Alexandra Scanameo and Nicholas P. Ziats
6.1 Introduction 95
6.2 Immune Responses to Decellularized Extracellular
Matrices 96
6.3 Innate Immunity 98