12.12.

2023

Stem Cells and Tissue Engineering

Tissue engineering
• Tissue Engineering evolved from the field of biomaterials
development and refers to the practice of
combining scaffolds, cells, and biologically active
molecules into functional tissues.

• The goal of tissue engineering is to assemble functional

constructs that restore, maintain, or improve damaged
tissues or whole organs.

• Artificial skin and cartilage are examples of engineered

tissues that have been approved by the FDA; however,
currently they have limited use in human patients.

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Regenerative Medicine

Helping the body to heal itself

• Tissue Engineering
• Cell Therapy
• Regeneration

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Artificial skin made from cowhide,

shark cartilage and plastic has
been successfully used by doctors
at the Massachusetts General
Hospital in Boston to replace skin
destroyed by burns.

Regenerative Medicine/Tissue Engineering

A field of research for over 70 years.

Why so few clinical advances?

• Inability to expand cells in vitro.

• Inadequate biomaterials
• Inadequate vascularity

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Inability to expand cells in vitro

Early 1990s-Most human cells could not be grown or expanded outside of the
body.

Inadequate Biomaterials

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Vascularity Problem
Cells alone can not survive in volumes greater than 0.3 mm 3

Nutrition to the cells is limited (limited vascularity)

Complex Organ Structures

Liver

Kidney

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Components of Tissue Engineering

• Cells
– Living part of tissue
– Produces protein and provides function of cells
– Gives tissue reparative properties

• Scaffold
– Provides structural support and shape to construct
– Provides place for cell attachment and growth
– Usually biodegradable and biocompatible

• Cell Signaling
– Signals that tell the cell what to do
– Proteins or Mechanical Stimulation

Components of Tissue Engineering

• Repair/replace damaged tissues
– Enhance natural regeneration

Cell Source
Embryonic stem cells
Tissue stem cells
Progenitor cells

Signals Scaffold
Growth factors Metals
Drugs Ceramics
Mechanical forces Synthetic polymers
Natural polymers

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Components of Tissue Engineering

Components of Tissue Engineering

Porous Scaffolds

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Important Variables
• Delivery
– Cell Suspensions Modify Cell
– Tissue-like constructs (scaffolds)
Behavior
• Chemical properties Survival
– Growth factors Organization
Migration
– Degradation particles Proliferation
– Matrix surface Differentiation
• Physical properties
– Structure
– Topography Optimize Cellular
– Rigidity Response
– Mechanical Loading

Key Chalenges

• Good sources: cells / scaffolds

• Complex tissues
• Vascularization
• In vivo functionality
• Cost efficient/scalable
• GMP/regulatory compliance

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Stem Cell Sources

• Autologous: Come from the person that needs

the new cells.
• Allogeneic: Come from a body from the same
species.
• Xenogenic: Come from a different species then
the organism they’re going into.
• Isogenic (Syngenic): Come from identical
twins.

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Purification

• Fluorescence-activated cell sorting (FACS)

• Magnetic Cell Sorting (MACS)
• Adhesion-Based Cell Sorting
• Complement Depletion
• Viral Labeling
• ……..

Overview Cell separation: https://app.jove.com/science-education/v/13371/overview-of-cell-separation-

and-isolation

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Video 1: FACS

Purification
Spesific surface marker

MACS

Video 2: MACS

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Scaffold Purpose

• Temporary structural support Structural

– Maintain shape
• Cellular microenvironment Surface
coating
– High surface area/volume
– ECM secretion
– Integrin expression
– Facilitate cell migration

Ideal Extracellular Matrix

• 3-dimensional
• Cross-linked
Modulate Properties
• Porous Physical, Chemical
Customize scaffold
• Biodegradable
• Proper surface chemistry
• Matching mechanical strength
• Biocompatible
• Promotes natural healing Appropriate Trade-offs
Tissue
• Accessibility Disease condition

• Proper architecture
• Commercial Feasibility

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Example scaffolds for bone tissue engineering

Carbohydrate Polymers 164 (2017) 200–213, Sevim Isik at al.

Delivery Methods

• Injectable stem cells

– Cells or cell-polymer mix
– Less invasive
– Adopt shape of environment
– Controlled growth factor release

• Solid scaffold manufacturing

– Computer-aided design
– Match defect shape

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“Natural” Materials
• Polymers
Perfusion-decellularized matrix: using nature's platform
– Collagen to engineer a bioartificial heart.
Ott, et al.
– Laminin Nat Med. 2008 Feb;14(2):213
– Fibrin
– Matrigel
– Decellularized matrix
• Ceramics
– Hydroxyapatite
– Calcium phosphate
– Bioglass

Materials for Tissue Engineering

• Acellular tissue matrix

Bioengineering of Organs

cadaveric rat hearts

Ott HC et al., Perfusion-decellularized matrix: using nature's platform to

engineer a bioartificial heart, Nature Medicine, 2008

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Decellularized matrix

WPI Team Grows Heart Tissue on Spinach Leaves

Researchers turn to the vascular system of plants to solve a major bioengineering problem
blocking the regeneration of human tissues and organs.
March 22, 2017

Video 2-Living skin substitutes

Challenge

To identify the best material, configuration

and coating is needed for optimal
construction of each target tissue

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Cell attachment and cell viability of hMSCs

on chitosan scaffolds

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This is an image of pancreatic islet cells encapsulated by an alginate bead. These beads are
rather large and contain much than just one cell per bead. The bead diameter can vary greatly
(from 25 micrometers to several millimeters).

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Regenerative Medicine Pioneers

Wake Forest Institute for Regenerative Medicine

(WFIRM)

Dr. Anthony
Atala

• 2006-2007: First to Engineer/Transplant Lab-Grown Organ

into a Human
• Transplant was Successful

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Regenerative Medicine Pioneers

Dr. Paolo Macchiarini (Karolinska Institute)

2008: Implanted World’s First Donor Trachea

• Recipient: Claudio Castillo
• Survived Procedure — Now Has Normal Respiratory
Function

Regenerative Medicine Pioneers

Dr. Ali Khademhosseini (Wyss Institute at Harvard)

[MIT Technology Review:ww2.technologyreview.com/tr35/profile.aspx?TRID=610]

• 2007: Creating living tissues
• Personalized 3D Vascular Constructs

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Regenerative Medicine Pioneers

McGowan Institute of Regenerative Medicine (University of Pittsburg)
Dr. Stephen Badylak

• Re-grows Severed Digits and

• New Muscle Tissue Development of 3-D
bioscaffolds for liver and heart regeneration

Regenerative Medicine Pioneers

Dr. Geraldine Hamilton (Wyss Institute, Harvard)

2011: Organ on a Chip Technology

(Drug Testing Tool)

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Potential of Regenerative
Medicine
• Chip Technology

[Geraldine Hamilton, Body parts on a chip, TEDx Boston, June 2013:

https://youtu.be/CpkXmtJOH84]

• Reduces Need for Animal Testing

• 3-D Printed Organs on Chips Used to Test Vaccines

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Regenerative Medicine Innovators

Dr. Jordan Miller (Rice University)

• 2013: Uses 3-D Print Technology
• Engineers Blood Vessels Using Sugar

Promise of Regenerative Medicine

• 3-D Printer Creates Heart Membrane

[Prof. Igor Efimov, Washington University in St. Louish:

ttps://news.wustl.edu/news/Pages/26554.aspx] Lizhi Xu and al., Nature
Communications, 2014, 3329, doi:10.1038/ncomms4329

3-D Elastic Membrane Fits Heart’s Epicardium

Video 3-Tissue engineering by 3D printers

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3D Printed Ear by Direct Ink Write (DIW) Technique

Futuristic!
Stem Cells + Organ Scaffold + 3D Printer

= Libraries of Replacement Organs?

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Related Video Links

https://www.jove.com/video/2561/isolation-and-culture-of-adult-epithelial-stem-cells-from-human-skin

https://www.jove.com/video/3194/isolation-culture-neural-crest-stem-cells-from-human-hair

https://www.youtube.com/watch?v=GqJYMgAcc0Q

•http://www.youtube.com/watch?v=F7WTe7_m76g

•http://www.youtube.com/watch?v=GwcT1ViM-hw&NR=1

•http://www.youtube.com/watch?v=0taE4F0Wkhg&feature=related

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