Tissue Engineering Resource Center
 

Training

Tissue Engineering Laboratory Course:
Offered during Fall Semester

Instructor:
Leah Abraham
Dept. Biomedical Engineering & Chemical & Biological Engineering
Phone: 617-627-2445
Email: leah.abraham@tufts.edu
Office Room 141, Science and Technology Center

Description:
The course is planned for upper level undergraduates and graduate students in the sciences and in engineering to gain a comprehensive understanding of the components involved in the field of tissue engineering, as well as the inter-related nature of these components in terms of functional tissue outcomes. For the hands-on laboratory studies, suitable background fundamentals and perspectives will be covered in handouts and via min-lectures. Basic laboratory safety, documentation, ethics and related topics will also be covered. Organization:

Module 1 - Scaffold Formation and Characterization
Module 2 - Cell Culture
Module 3 - Bioreactors and Integration

Module 1 - Scaffold Formation and Characterization
Introduction - This module will give the students an opportunity to learn how to design, prepare and characterize scaffolds that are needed for tissue engineering research. The students will be given a brief introduction to the properties and applications of various biomaterials with emphasis on biopolymers like collagen, silk and polylactic acid. The students will learn how to design scaffolds with consideration for cell biology, mechanics, degradation, biocompatibility and clinical relevance. Methods to characterize the scaffolds through microscopic and analytical tools will be covered. The focus will be the design and preparation of 2D and 3D scaffolds with appropriate surface and bulk properties (structure, morphology, mechanics) for various engineered tissue types. The students will also be introduced to the formation of spongy 3D matrices and to electrospinning for formation of fibers.

Chap 1.Scaffolds and Biomaterials
1.1.Introduction
1.2.Time-table plan
1.3.2D Films and 3D Scaffolds
1.3.1.Collagen Film Protocol
1.3.2.Silk Scaffold Protocol
1.3.3.Protocol for assay
1.4.Electrospinning
1.4.1.Protocol of Electrospinning of Regenerated Silk Fibroin
1.5.Protocol Solution
1.6.Additional Reading

Module 2 - Cells and Cell Culture
Introduction - Background on basic cell culture techniques including aseptic skills, cell expansion, cell transfer, cell storage and cell characterization will be covered. The focus will be on human bone marrow derived mesenchymal stem cells.

Chap 2.Introduction to Cells and Cell Aseptic Skills
2.1.Introduction
2.2.Cells
2.2.1.Primary cells
2.2.2.Secondary cells
2.2.3.Immortalized cells
2.3.Cultured Cells
2.4.Cell types
2.5.Source of cells
2.6.Cell culture systems
2.7.Human mesenchymal stem cells
Chap 3.Aseptic Techniques
3.1.General Guidelines
3.1.1.Protocol for using a hood
3.1.2.CO2 incubators
3.1.3.Microscopes
3.2.Sterile Conditions
Chap 4.Cell Culture Media
4.1.Types and composition
4.2.Media Preparation
4.3.Other Supplements and Additives
4.3.1.Serum
4.3.2.Microscopes
4.3.3.Vessels
4.3.4.Safety consideration
Chap 5.Tissue culture practice
5.1.Adherent cell cultures
5.1.1.Initiating a cell culture
5.1.2.Resuscitation of frozen cells
5.1.3.Maintaining a cell culture
5.1.4.Harvesting cells
5.1.5.Cell counting
5.1.6.Freezing cells
5.2.Semi-adherent cell culture
5.3.Suspension cell culture
5.4.Surface markers
5.5.Module 2 plan for cell culture
5.6.Additional Reading

Module 3 - Bioreactors and Integration
Introduction - Students will learn principles of bioreactor design for in vitro tissue engineering. Both static and dynamic bioreactors will be included. Emphasis will be on bioreactors for engineering of skeletal tissue such as bone, cartilage, ligaments and myocardial. Aspects of mechanics, transport, scale and imaging will be included in the material. Students will learn how to integrate the previous three modules into a specific system for functional tissue engineering. The students will learn this through three case studies in vitro involving bone, cartilage, myocardial and ligament tissue engineering. The cells and scaffolds prepared in the previous modules will be used in these case studies. The student will also learn how to evaluate the outcomes of the engineered tissues through various biological, chemical and mechanical testing. A number of tissue specific assays will be used to assist the process. At the end of this module, the students will have established an overall systems strategy to functional tissue engineering in vitro. As a final assignment, they will be responsible for the design of a new system to use towards engineering a new type of tissue for functional relevance towards clinical application, or for in vitro study of disease models or therapeutic strategies.

Chap 6.Bioreactors and Integration
6.1.Introduction
6.2.Types of Reactors
6.2.1.Spinner Flask
6.2.2.Rotating vessel
6.2.3.Perfused Column and Perfused Chamber
6.3.Plan for module 3: Bioreactors and Integration
Chap 7.Medium preparation
7.1.Basic media
7.2.Osteogenic Media
7.3.Adipogenic media
Chap 8.MSC seeding on 2D films and 3D scaffolds
8.1.MSC seeding on 3D polymer scaffolds
8.2.MSC static and dynamic cultivation on 3-D polymers
8.3.Adipogenic differentiation on 2D denatured collagen film
Chap 14.Assays
14.1MTT analysis
14.2Oil Red-O Staining and Quantification of Lipids
14.3Hematoxylin and Eosin
14.4Additional Reading

** This description is from the first session in which the class was taught. Based on feedback from that group there may be some minor changes to the above content.

Tissue Engineering Laboratory Course:
Offered during Summer Semester

Instructor:
David Kaplan
Professor, Dept. Biomedical Engineering & Chemical & Biological Engineering
Director, Tufts University Bioengineering & Biotechnology Center
Phone: 617-627-3251
Email: david.kaplan@tufts.edu
Office Room 153, Science and Technology Center

Description:
The course is planned for upper level undergraduates and graduate students in the sciences and in engineering to gain a comprehensive understanding of the components involved in the field of tissue engineering, as well as the inter-related nature of these components in terms of functional tissue outcomes. For the hands-on laboratory studies, suitable background fundamentals and perspectives will be covered in handouts and via min-lectures. Basic laboratory safety, documentation, ethics and related topics will also be covered.

Organization:
Module 1 - Scaffold Formation and Characterization
Module 2 - Cell Culture
Module 3 - Bioreactors and Integration

Module 1 - Scaffold Formation and Characterization

Introduction - This module will give the students an opportunity to learn how to design, prepare and characterize scaffolds that are needed for tissue engineering research. The students will be given a brief introduction to the properties and applications of various biomaterials with emphasis on biopolymers like collagen, silk and polylactic acid. The students will learn how to design scaffolds with consideration for cell biology, mechanics, degradation, biocompatibility and clinical relevance. Methods to characterize the scaffolds through microscopic and analytical tools will be covered. The focus will be the design and preparation of 2D and 3D scaffolds with appropriate surface and bulk properties (structure, morphology, mechanics) for various engineered tissue types. The students will also be introduced to the formation of spongy 3D matrices and to electrospinning for formation of fibers.

Chap 1.Scaffolds and Biomaterials
1.1.Introduction
1.2.Time-table plan
1.3.2D Films and 3D Scaffolds
1.3.1.Collagen Film Protocol
1.3.2.Silk Scaffold Protocol
1.3.3.Protocol for assay
1.4.Electrospinning
1.4.1.Protocol of Electrospinning of Regenerated Silk Fibroin
1.5.Protocol Solution
1.6.Additional Reading

Module 2 - Cells and Cell Culture

Introduction - Background on basic cell culture techniques including aseptic skills, cell expansion, cell transfer, cell storage and cell characterization will be covered. The focus will be on human bone marrow derived mesenchymal stem cells.

Chap 2.Introduction to Cells and Cell Aseptic Skills
2.1.Introduction
2.2.Cells
2.2.1.Primary cells
2.2.2.Secondary cells
2.2.3.Immortalized cells
2.3.Cultured Cells
2.4.Cell types
2.5.Source of cells
2.6.Cell culture systems
2.7.Human mesenchymal stem cells
Chap 3.Aseptic Techniques
3.1.General Guidelines
3.1.1.Protocol for using a hood
3.1.2.CO2 incubators
3.1.3.Microscopes
3.2.Sterile Conditions
Chap 4.Cell Culture Media
4.1.Types and composition
4.2.Media Preparation
4.3.Other Supplements and Additives
4.3.1.Serum
4.3.2.Microscopes
4.3.3.Vessels
4.3.4.Safety consideration
Chap 5.Tissue culture practice
5.1.Adherent cell cultures
5.1.1.Initiating a cell culture
5.1.2.Resuscitation of frozen cells
5.1.3.Maintaining a cell culture
5.1.4.Harvesting cells
5.1.5.Cell counting
5.1.6.Freezing cells
5.2.Semi-adherent cell culture
5.3.Suspension cell culture
5.4.Surface markers
5.5.Module 2 plan for cell culture
5.6.Additional Reading

Module 3 - Bioreactors and Integration

Introduction - Students will learn principles of bioreactor design for in vitro tissue engineering. Both static and dynamic bioreactors will be included. Emphasis will be on bioreactors for engineering of skeletal tissue such as bone, cartilage, ligaments and myocardial. Aspects of mechanics, transport, scale and imaging will be included in the material. Students will learn how to integrate the previous three modules into a specific system for functional tissue engineering. The students will learn this through three case studies in vitro involving bone, cartilage, myocardial and ligament tissue engineering. The cells and scaffolds prepared in the previous modules will be used in these case studies. The student will also learn how to evaluate the outcomes of the engineered tissues through various biological, chemical and mechanical testing. A number of tissue specific assays will be used to assist the process. At the end of this module, the students will have established an overall systems strategy to functional tissue engineering in vitro. As a final assignment, they will be responsible for the design of a new system to use towards engineering a new type of tissue for functional relevance towards clinical application, or for in vitro study of disease models or therapeutic strategies

Chap 6.Bioreactors and Integration
6.1.Introduction
6.2.Types of Reactors
6.2.1.Spinner Flask
6.2.2.Rotating vessel
6.2.3.Perfused Column and Perfused Chamber
6.3.Plan for module 3: Bioreactors and Integration
Chap 7.Medium preparation
7.1.Basic media
7.2.Osteogenic Media
7.3.Adipogenic media
Chap 8.MSC seeding on 2D films and 3D scaffolds
8.1.MSC seeding on 3D polymer scaffolds
8.2.MSC static and dynamic cultivation on 3-D polymers
8.3.Adipogenic differentiation on 2D denatured collagen film
Chap 14.Assays
14.1MTT analysis
14.2Oil Red-O Staining and Quantification of Lipids
14.3Hematoxylin and Eosin
14.4Additional Reading


 
 
 
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