3D Cell Cultures: Technologies and Global Markets

Table of Contents

Chapter 1 Introduction

• Study Goals and Objectives
• Reasons for Doing the Study
• Scope of Report
• Intended Audience
• Methodology and Information Sources
• Analyst's Credentials
• Custom Research
• Related Reports

Chapter 2 Summary

• New Applications Identified Since 2015 Report
• Changes from 2015 Report
• Changes from 2017 Report
• New Applications Since 2017
• Changes Since 2017

Chapter 3 Highlights and Issues

• An Opening Comment on an Amazing Industry
• Industry Issues
• In Vitro versus In Vivo
• Dimensionality
• The Research Chain for 2D and 3D Cell Culture
• Best Practices
• Standardization
• Regulation
• Genomics Forcing the Hand of the FDA
• Leachables and Extractables
• Broad Issues
• Research Talent Shortages
• The Shifting International Picture
• Pace and Diversification of Innovation
• A Comment on the "Other" Areas of Cell Culture
• Omics Everywhere
• Is 2020 a Watershed Year for the Cell Culture Industry?
• Preliminary Market Analysis
• Cell Culture Market Growth Rate Estimates
• Assessing Large-Scale Media Consumption Needs
• Modeling Future Growth in Biopharmaceuticals
• Base Case for the Cell Culture Market
• Challenges in Projecting Sales and Growth
• Cell Culture Media Market Estimates
• Cell and Gene Therapy Bioprocessing Segment
• Evaluating Media Consumption for Biosimilars
• What About CDMOs?
• Microfluidics
• Bioreactors
• Internal Cell Culture Resources
• Drug Development Costs: What to Believe?
• Is There Too Much Concentration of Ownership in Biotechnology?
• Characterizing Innovation in 3D Cell Culture
• Bioprinting Strategic Roadmap
• Impact of COVID-19 on the World of Cell Culture
• Clinical Trials on COVID-19

Chapter 4 Tissue and Cell Culture: Technology and Product Background

• Where did Tissue and Cell Culture Start?
• History and Early Applications
• Invention of Tissue Culture
• Development of Sustained Cell Lines
• First Cell Culture Flask and Rigorous Techniques
• Lindbergh: The Cell Culture Equipment Pioneer
• Establishing Continuous Cell Lines
• Key Developments in Equipment
• Terminology and Concepts
• Tissue and Cell Culture Industry
• Tissue
• Cells
• Cell Types
• Tissue Culture and Cell Culture Definitions
• Cell Lines
• Care and Growth of Cell Culture Systems
• Consumables
• Media, Sera and Reagents
• Gels and Scaffolds
• Microplates/Microtiter Plates
• Equipment
• Bioanalytical Instruments
• Bioanalytical Imaging
• Bioprinting
• Bioreactors
• Other Equipment for Cell Culture
• Adherent Approaches
• Traditional Roller Bottles
• Other Systems
• Information Technology: Software and Services for the Cell Culture Research Market
• Software for the Research Market in Cell Culture
• Software-Related Support Services
• Bioprocessing Consumables for Cell Culture
• Microcarriers for Large-Scale 3D Culture
• Sera for Large-Scale 3D Culture
• Media for Large-Scale 3D Culture
• Bioreactor Bags for Large-Scale 3D Culture
• Other
• Bioprocessing Equipment
• Analytical Equipment for Bioprocessing
• Automation Systems for Bioprocessing
• Support Equipment for Bioprocessing
• Aspects of Large-Scale Manufacturing of Biopharmaceuticals and Vaccines
• Suspension Proteins and Monoclonal Antibodies
• Adherent-Cell-Based Therapies and Vaccines
• Small-Scale Adherent to Make Somatic Cells, Stem Cells and Tissues
• Vaccines
• Vaccine Development as a Catalyst
• Vaccines Developed Using Human Cell Strains
• Exosome Manufacturing
• Viral Vector Manufacturing
• Lentivirus Manufacturing
• Plasmid Manufacturing
• Cell Culture End Users
• Pharma/Biopharma
• Universities
• Government
• CROs/CDMOS
• Other
• Cell Culture Applications
• Drug Discovery
• Clinical Development
• Toxicology
• Basic Research
• Bioprocessing Development
• Other
• Regional Markets
• The Americas
• Europe
• Asia-Pacific
• Rest of the World

Chapter 5 Assays, Imaging and Analysis

• Assays
• Assay Development for Mesenchymal Stem Cells
• In Vitro Testing of Adventitious Agents
• Assays and Assay Kits
• Cell-Based Assays: Overview and Newer Developments
• Cells Used in Cell-Based Assays
• Notes on 3D Cell-Based Assays
• Kinetic Metabolism Assays
• Cell Proliferation
• Viability and Cytotoxicity
• Permeability Assays for Cell Viability and Survival
• Cell Invasion
• Cell Signaling and Communication
• Cytostatic
• Cell Death Assays
• Imaging Technology
• Imaging Assays
• Fluorescence as a Driver of Screening
• Analytical Systems Used in Tissue and Cell Culture
• Understanding ""Cellomics""
• HCS Support of 3D Cell Culture
• NGS Discovery Pools
• Multiplex Assays
• Predictive Toxicology
• Neuro Safety
• The Omics Invasion
• Transcriptomics

Chapter 6 Regulation and Standardization

• U.S. Regulatory Status of Bioprinted Products
• Basic Guidance for the Regulation of Biologics
• Guidance for Regenerative Medicine
• Guidance for Xenotransplants
• Guidance for Regenerative Medicine: Emergency Approval
• Regulating Bioprinted Products

Chapter 7 3D Models for Cancer

• Disease Modeling
• Cancer
• Main Classes of Models for Researching Cancer and Other Diseases
• Cell Lines
• Spheroids and Organoids
• Genetically Engineered Mouse Model (GEMM)
• Patient-Derived Tumor Xenografts (PDXs)
• Overview: Cancer at the Cellular Level
• In Vivo (Animal) Testing Standard
• Empire of the Mouse
• Humanized Mice
• 2D Culture
• 3D Requirements
• Cell Number and Viability
• Migration and Invasion
• Unmet Needs: Angiogenesis and Immune System Evasion
• Benefits of 3D Models to Cancer Research
• Greater Distinction in Cell Morphology and Proliferation
• Greater Gene Expression and Cell Behavior
• Better Models of Cell Migration and Invasion
• Cell Heterogeneity
• Breast Cancer as a Driver of 3D Cultures
• Structure, Polarity and Apoptosis
• Melanoma as a Driver of 3D Cultures
• Moving to Spheroid Configurations
• 3D Systems in Cancer Research
• Multicellular Tumor Spheroids
• Multilayered Cell Cultures
• 3D Engineered Scaffolds
• Natural Materials
• Synthetic Materials
• Human Cancer Model Initiative (HCMI)
• Next-Generation Human Cancer Models
• Drug Sensitivity and Resistance
• Altered Signaling and Sensitivity
• Drug Resistance
• Cellular Signaling
• Cellular Signaling Mediated by Integrins
• Drug Screening
• Approaches and Endpoints
• Spheroid Applications
• Metastasis via 3D Cell Migration Model
• Metastasis via Lung-on-Chip
• Cancer Metabolism
• Future Horizons
• Metastases
• Coculture
• Vascularization
• Cancer-Associated Fibroblasts
• Cancer Stem Cells
• Combination Therapies
• Biologics Development
• Tumor Recurrence
• Patient-Derived Cells
• Patient-Derived Tumor Xenografts (PDXs)
• Evolution of PDX Platforms

Chapter 8 Landscape for Toxicology and Drug Safety Testing

• Introduction
• Liver
• Cardiovascular Toxicity
• Toxicology Background
• Testing for Adverse Effects on the Skin
• New Assessment Methodologies Impact on 3D Cell Culture
• Toxicology Testing in Cosmetics
• Updated Regulatory Requirements
• Efficacy of Cosmetics and Cosmeceuticals
• Aspects of Cosmetic Toxicity Testing
• Skin Irritation
• Skin Corrosion
• Phototoxicity
• Skin Sensitization
• Eye Irritation
• Acute Systemic Toxicity
• Acute Toxicity Testing
• Cytotoxicity Assays for Acute Toxicity Testing
• Chronic and Repeated Dose Toxicity
• Carcinogenicity and Genotoxicity
• Overview
• In Vitro Methods: Background and Recent Developments
• Regulatory versus Drug Development Applications
• Efforts to Reduce False Positives
• Recent Innovations in Screening
• Future Challenge: Non-genotoxic Carcinogens
• Reproductive and Developmental Toxicity
• Background
• Following the Reproductive Cycle
• Development and Reproductive Tox Testing Types
• Zebrafish Model for Developmental Toxicity Screening
• Combination of Zebrafish and Stem Cells
• Biomedical Frontiers: Male Testis
• Endocrine Disruptor Screening
• Background
• Environmental Toxicology Impacts In Vitro Methods
• High-Production Volume Chemicals
• ToxCast and Tox 21 Initiatives
• Future Challenge: Thyroid Disruption
• BG1 Assay
• Toxicokinetics and ADME
• In Vitro Developments
• Metabolism
• Pharmacokinetics of Low Turnover Compounds
• Organotypic Models
• 3D Models for Skin
• 3D Corneal System
• Absorption Barrier Models
• Gastrointestinal
• Lung
• Blood-Brain Barrier
• Real Architecture for 3D Tissue Barriers and Extracellular Matrix
• Liver Toxicity
• Uniqueness and Complexity of Liver
• Liver as a Key Driver for 3D Innovation
• In Vitro Liver Applications
• In Vivo Liver Function and Structure
• Liver Metabolism
• In Vitro Liver Models
• Cocultures of Hepatocytes and Macrophages
• 3D Liver Models
• Bioprinted Liver Tissue
• Detected Hepatosphere Structures and Functionality
• Ideal Criteria
• Drug Resistance
• Transporter Studies
• Achieving Heterotypic Cell-Cell Contacts
• Unmet Needs and Future Drivers of Innovation
• Morphogen Signaling
• Multi-donor Liver Cells
• Kidney Toxicity Applications
• Future Challenge: Stem-Cell Derived Kidney Cells
• Bioprinted Kidney Tissue
• Pancreatic Toxicology Applications
• Cardiovascular Toxicity
• Commercial Avenue
• Collaboration with Regulators
• Cardiovascular Drug Discovery
• Microelectrode Arrays (MEAs) Based on Impedance
• Surrogate for Aortic Ring Assay via Bioprinted Magnetics
• Vasodilator Activity
• 3D Engineered Heart Tissues

Chapter 9 Stem Cell Landscape

• A History of Stem Cells
• Major and Minor Research Areas for Stem Cells
• 3D Stem Cell Culture Systems
• Plate or Culture Dish
• Spinner Flask and Rotating Wall Vessel
• Perfusion Bioreactor and Microcarrier Systems
• Shortcomings
• Microfluidics and Stem Cells
• Short Review of Stem Cell Biology
• Embryogenesis
• Growth, Structure and Morphology of Stem Cells
• Stem Cell Differentiation
• Stem Cell Differentiation versus Proliferation
• Extracellular Matrix and Stem Cells
• Soluble Factors
• Manufacturing Stem Cells
• Controlling Embryoid Body Formation
• Forced Aggregation Cultures
• Hanging-Drop Approach
• Applications
• Stem Cell Markers for High-Throughput Screening
• Teratomas and the Teratoma Assay
• Fujifilm (Cellular Dynamics Inc.)
• Stem Cell Research Breakthroughs
• Stem Cells for Neuroscience Discovery and Development
• Example: Alzheimer's Research
• Background: B-Amyloid Cascade Hypothesis
• Human iPSC-Derived Models
• New 3D Model
• Other Advantages of 3D
• Envisioned Applications
• Other 3D Neuro Applications
• Stem Cells for Cardiovascular Discovery
• Stem Cells for the Development of Regenerative Medicine
• Background: Allogenic versus Autologous
• MicroRNAs
• Induced Pluripotent Stem Cells (IPS)

Chapter 10 Regenerative Medicine: Organ Transplants and Skin Substitutes

• Regenerative Medicine
• Need for Organ Transplants
• Applications in Regenerative Medicine
• Investments in Regenerative Medicine
• Skin Substitutes Industry
• Tissue Culture Allograft and Autograft Products
• Tissue Engineering in Regenerative Medicine

Chapter 11 Company Profiles

• 3D Biopinting Solutions
• 3D Biotek Llc
• 4D Technology
• Abcam
• Akron Biotechnology Llc
• Amsbio Llc
• Agilent Technologies
• Alpco
• Applikon Biotechnology
• Beckman Coulter Life Sciences
• Biogelx Ltd.
• Bioinspired Solutions
• Biotime Inc.
• Biovision Inc.
• Cell Applications Inc.
• Cellink
• Cellspring
• Corning Life Sciences
• Cyprotex Plc
• Cytiva
• Cytoo Sa
• Emd Millipore
• Emulate Inc.
• Envisiontec Inc.
• Epithelix Sarl
• Eurofins Sas
• Greiner Bio-One
• Hamilton Robotics
• Hubrecht Organoid Technology
• Hurel Corp.
• Insphero Ag
• Invitrocue Ltd.
• Kiyatec Inc.
• Lifenet Health
• Lorem Cytori
• Mattek Corp.
• Mimetas Inc.
• Mitra Biotech Inc.
• N3D Biosciences Inc.
• National Center For Advancing Translational Sciences (Ncats)
• Organovo
• Pandorum Technologies
• Perkinelmer
• Plasticell Ltd.
• Pluristem Therapeutics, Inc.
• Poietis Sas
• Promega Corp.
• Qualyst Transporter Solutions Llc (Subsidiary Of Bioivt)
• Regenhu Ltd.
• Regenovo Biotechnology
• Screen Holdings Co. Ltd.
• Seahorse Bioscience
• Stemcell Technologies Inc.
• Stratatech Corp.
• Synvivo Inc.
• Tap Biosystems
• Tecan Group Ltd.
• Tevido Biodevices
• Thermo Fisher Scientific
• Zen-Bio Inc.

List of Tables
Summary Table: Global Market for Cell Culture, by Segment, Through 2025
Table 1: Dimensionality of Cell Culture
Table 2: WHO R&D Roadmap of Priority Infectious Diseases
Table 3: 3D Bioprinting Roadmap
Table 4: Selected Online Prices for Bioreactors
Table 5: Current Clinical Trials on Remdesivir
Table 6: Tissue Types
Table 7: Cell Types Based on Developmental Origin
Table 8: Leading Cell Line Suppliers, April 2020
Table 9: Commonly Used Transformed Cells Lines
Table 10: Major Primary Cell Lines
Table 11: Leading Primary Cell Suppliers, April 2020
Table 12: Main Types of Stem Cells
Table 13: Stem Cell Services
Table 14: Areas of Interest in 3D Spheroid Research
Table 15: Global Market for Cell Culture Consumables in Research, Through 2025
Table 16: Selected Nanoparticle Products Used in Life Science Research
Table 17: Results of Liver-Chip Drugs Halted in Previous Clinical Trials Based on Animal Studies
Table 18: Companies and Universities Involved in the Organ-on-a-Chip Industry
Table 19: Global Market for Cell Culture Equipment, by Type, Through 2025
Table 20: Microfluidics Companies
Table 21: Selected Recent Patents Issued on Microfluidic Devices Related to Cell Culture Applications
Table 22: Selected Papers Published on Recent Microfluidic Advances in Cell Culture
Table 23: High-Content Screening Suppliers and Key Attributes, April 2020
Table 24: Selected Patents Issued Related to Flow Cytometers
Table 25: Thermo Fisher Imaging Products
Table 26: Publicly Disclosed Funding of Bioprinting Companies, 2017-2020*
Table 27: Bioprinting Company Deals and Strategic Partnerships, 2019 and 2020
Table 28: Bioprinting Modalities
Table 29: Maintaining Cell Viability During Printing
Table 30: Bioprinting Instrument Industry
Table 31: Biomaterial Components
Table 32: Bioink Types
Table 33: Classes of Matrix Bioink Hydrogels
Table 34: Matrix Bioink Selection Criteria
Table 35: Selected Bioink Companies, 2020
Table 36: Projected Unit Sales of Research Bioreactors, <10 Liters, 2020
Table 37: Leading Bioreactor Suppliers, 2020
Table 38: Label-Free Technologies and Suppliers, April 2020
Table 39: Global Market for Software and Services for Cell Culture Research, Through 2025
Table 40: Global Market for Bioprocessing Consumables for Cell Culture, Through 2025
Table 41: Global Market for Cell Culture Bioprocessing Equipment, by Type, Through 2025
Table 42: Global Market for Software for Cell Culture in Bioprocessing, Through 2025
Table 43: Developmental Issues Facing the Commercialization of Exosomes
Table 44: Companies Working on Exosome Products
Table 45: Global Market for Cell Culture, by End User, Through 2025
Table 46: Global Market for Cell Culture, by Application, Through 2025
Table 47: Global Market for Cell Culture, by Region, Through 2025
Table 48: Typical Assay Endpoints and Tests
Table 49: U.S. Patents on Assays Systems, 2019 and 2020
Table 50: Selected U.S. Patents on Assay Imaging, 2019 and 2020
Table 51: Recently Issued U.S. Patents on Cellomics, 2017-2019
Table 52: Toxicology Issues That Need to Be Addressed for FDA-Regulated Products
Table 53: Overview of Federal Regulation of the Cell Culture Markets
Table 54: FDA List of Cell, Biologic and Tissue Products Regulated Under CBER and CDRH
Table 55: FDA “Talking Point” Recommendations for Regenerative Medicine Advanced Therapies (RMATs)
Table 56: U.S. Regulatory Considerations
Table 57: Main Types of Models for Researching Cancer and Other Diseases
Table 58: Goals of Funding Opportunity Announcement RFA-CA-19-055
Table 59: Common 3D Assays
Table 60: 3D Tumor Models Developed for Drug Discovery, June 2017
Table 61: Summary of the Benefits and Advantages of EV3D
Table 62: ECVAM List of Current Activities, 2020
Table 63: In Vitro Testing in Cosmetics, by Test Class
Table 64: In Vitro and In Vivo Assays in the Tier 1 Screening Battery, January 2014
Table 65: Timeline of Key Developments in Stem Cell Research, 1978-2006
Table 66: Major and Minor Research Areas in Stem Cells
Table 67: Stem Cell Usage in Research
Table 68: Recent U.S. Patents Granted on Stem Cell Technologies
Table 69: Papers Published on Selected Stem Cell Research Trends
Table 70: Comparison of the Number of Citations in the Literature on Cell Culture versus Stem Cell Culture
Table 71: Number of Organ Transplants Performed in the United States, 2018 and 2019
Table 72: Total Global Financing of Regenerative Medicine
Table 73: Total Financing of Regenerative Medicine, by Therapeutic Area
Table 74: Leading Tissue Products and Suppliers, April 2020
Table 75: Printed Tissue and Organs: Commercialization Timeframe
Table 76: Tissue/Organ Complexity
Table 77: New Bioassay Kits Introduced by BioVision
Table 78: Partial List of Specialty Media Formulations Sold by Cell Applications Inc.

List of Figures
Summary Figure: Global Market for Cell Culture, by Segment, 2020-2025
Figure 1: Research Chain for Cell Culture
Figure 2: A Model for The Evolution of FDA Regulation
Figure 3: An Innovation Matrix
Figure 4: Bioprinting Strategic Roadmap
Figure 5: Gastrointestinal Organotype Cultures
Figure 6: iCELLis Nanoreactor: Example of Commercial 2D Cell Culture Systems
Figure 7: Prototype of 3D Model Lung-on-a-Chip from Wake Forest
Figure 8: Recent Photos of the HepaChip
Figure 9: Recent Photos of the HepaChip-MWP
Figure 10: University of Toronto Handheld Bioprinting Device
Figure 11: Collaborative Experiment Conducted by American, Russian and Israeli Scientists
Figure 12: Smart Marbles Concept for Quantifying Process Heterogeneity
Figure 13: Porcine Intestinal Organoids
Figure 14: Diagram of the Components of a Predictive Toxicology System
Figure 15: Gleason’s Pattern
Figure 16: Diagram of PDXs, Cell Lines and Organoid/Spheroid Xenografts
Figure 17: Cell Heterogeneity and Its Function
Figure 18: Electron Micrograph of Porous Microcarrier for Stem Cell Production
Figure 19: Photograph of Apligraf

Executive Summary

Reasons for Doing the Study:

Three-dimensional cell culture is a key area for needed progress in research and commercialization in biotechnology. Where cell culture is a cornerstone of biotechnology, the 3D component has reinforced its role in strengthening other parts of biotechnology. Whether the discussion is about stem cells, tissue engineering, or microphysiological systems, their vital role in drug discovery, toxicology, and other areas leading to new product development, 3D cell culture is becoming the environment that will increasingly define the basis for future advances.

To mix metaphors, 3D cell culture is also cross-roads through which just about everything else passes on its way to building knowledgebases or introducing new products. This study is needed to bring together and make sense out of the broad body of information encompassed by 3D cell culture.

Three-dimensional cell culture has been used by researchers for many years now, with early adoption and now key roles in cancer and stem cells. Organ-on-a-chip technology, also known as microphysiological systems, is leading to dramatic breakthroughs. Also, stem cell research coupled with synthetic biology is opening new areas. This study is needed to provide a perspective on these advances. Furthermore, classical toxicology testing programs have been in place for many decades, and over the past 20 years, animal welfare and scientific activities have spurred the development of in vitro testing methods. In silico methods are advancing in novel ways that need to be analyzed and considered in terms of their impacts on cell culture.

This report investigates the recent key technical advances in 3D cell culture equipment, raw materials, assay kits, analytical methods, and clinical research organization (CRO) services. It should also be pointed out that this report takes a somewhat different position on 2D cell culture. It has been criticized for its inadequacies and the misleading information it can produce. However, a review of industry practices makes it clear that it still has its place and will contribute to future advances in unexpected ways.

The company section looks at many of the suppliers who provide equipment, assays, cells, reagents, and services used in 3D cell culture. This study sought to understand business models and market maturity dynamics in greater depth as well as providing more quantitative analysis of their operations.

Companies Mentioned

• 3D Biopinting Solutions
• 3D Biotek Llc
• 4D Technology
• Abcam
• Agilent Technologies
• Akron Biotechnology Llc
• Alpco
• Amsbio Llc
• Applikon Biotechnology
• Beckman Coulter Life Sciences
• Biogelx Ltd.
• Bioinspired Solutions
• Biotime Inc.
• Biovision Inc.
• Cell Applications Inc.
• Cellink
• Cellspring
• Corning Life Sciences
• Cyprotex Plc
• Cytiva
• Cytoo SA
• EMD Millipore
• Emulate Inc.
• Envisiontec Inc.
• Epithelix Sarl
• Eurofins Sas
• Fujifilm (Cellular Dynamics Inc.)
• Greiner Bio-One
• Hamilton Robotics
• Hubrecht Organoid Technology
• Hurel Corp.
• Insphero Ag
• Invitrocue Ltd.
• Kiyatec Inc.
• Lifenet Health
• Lorem Cytori
• Mattek Corp.
• Mimetas Inc.
• Mitra Biotech Inc.
• N3D Biosciences Inc.
• National Center For Advancing Translational Sciences (Ncats)
• Organovo
• Pandorum Technologies
• Perkinelmer
• Plasticell Ltd.
• Pluristem Therapeutics, Inc.
• Poietis Sas
• Promega Corp.
• Qualyst Transporter Solutions Llc (Subsidiary Of Bioivt)
• Regenhu Ltd.
• Regenovo Biotechnology
• Screen Holdings Co. Ltd.
• Seahorse Bioscience
• Stemcell Technologies Inc.
• Stratatech Corp.
• Synvivo Inc.
• Tap Biosystems
• Tecan Group Ltd.
• Tevido Biodevices
• Thermo Fisher Scientific
• Zen-Bio Inc.