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Methods in Bioengineering: Cell Transplantation Unabridged edition [Hardback]

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  • Formāts: Hardback, 272 pages
  • Izdošanas datums: 31-May-2011
  • Izdevniecība: Artech House Publishers
  • ISBN-10: 1608070158
  • ISBN-13: 9781608070152
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Methods in Bioengineering: Cell Transplantation Unabridged editionPalielināt
  • Formāts: Hardback, 272 pages
  • Izdošanas datums: 31-May-2011
  • Izdevniecība: Artech House Publishers
  • ISBN-10: 1608070158
  • ISBN-13: 9781608070152
Citas grāmatas par šo tēmu:
Permanent link: https://www.kriso.lv/db/9781608070152.html
Keywords:
This practical book is part of the new Artech House Methods in Bioengineering series - volumes designed to offer detailed guidance on authoritative methods for addressing specific bioengineering laboratory challenges. Written and edited by recognized experts in the field, each book provides research engineers, scientists, and students with step-by-step procedures, clear examples, and effective ways to overcome possible problems. This unique volume covers in detail the basic techniques and protocols of regenerative medicine, focusing on methods for cell transplantation. Readers explore a wide range of critical factors, such as grown tissues, cells, laboratory-made compounds, and engineered organs. The books helps professionals understand the many intricate steps involved in tissue engineering, cell isolation, progenitors and stem cell differentiation, and in vivo models for assessing new strategies.
Preface xvii
Chapter 1 Extracellular Matrices for Hepatic Tissue Engineering
1(22)
1.1 Introduction
2(1)
1.2 Experimental Design
2(1)
1.3 Materials
3(1)
1.4 Methods
3(15)
1.4.1 Cell Adhesion Assay
3(1)
1.4.2 Cell Detachment Assay
4(1)
1.4.3 Inhibition Assay
4(2)
1.4.4 Western Blot Assay
6(1)
1.4.5 Flow Cytometry Analysis
7(1)
1.4.6 Lectin Binding Assay
7(1)
1.4.7 Coupling of Galactose Groups with ECM Bulk and ECM Surface
8(3)
1.4.8 Regulation of Density of Galactose and Spatial Orientation of Galactose
11(3)
1.4.9 Regulation of Topology of ECM
14(1)
1.4.10 Regulation of Deposition of Natural ECM
15(1)
1.4.11 Design of Coculture System
15(2)
1.4.12 Choice of Cell Sources
17(1)
1.5 Anticipated Results
18(1)
1.6 Discussion and Commentary
18(1)
1.7 Application Notes
19(1)
1.8 Summary Points
19(4)
Acknowledgments
20(1)
References
20(3)
Chapter 2 Tissue Engineering Using a Temperature-Responsive Cell Culture Dish
23(12)
2.1 Introduction
24(1)
2.2 Experimental Design
24(1)
2.3 Materials
24(3)
2.4 Methods
27(2)
2.4.1 Seeding Cells
27(1)
2.4.2 Change Medium
28(1)
2.4.3 Harvest of Cell Sheets
28(1)
2.4.4 Transplantation of Cell Sheets
28(1)
2.4.5 Multilayering of Cell Sheets
28(1)
2.5 Data Acquisition, Anticipated Results, and Interpretation
29(1)
2.5.1 Autograft/Allograft Models
29(1)
2.5.2 Xenograft Models
29(1)
2.6 Discussion and Commentary
29(1)
2.7 Applications Notes
30(1)
2.7.1 Single Cells
30(1)
2.7.2 Cell Sheets
30(1)
2.8 Summary Points
31(4)
Acknowledgments
31(1)
References
32(3)
Chapter 3 In-Vivo Functional Assessment of Engineered Human Insulin-Producing Cells
35(12)
3.1 Introduction
36(1)
3.2 Experimental Design
37(1)
3.3 Materials
38(1)
3.4 Methods
38(4)
3.4.1 Generation of Hyperglycemic Immunodeficient Recipient Mice
38(1)
3.4.2 Cell Transplantation Under the Renal Capsule
39(1)
3.4.3 Monitoring Blood Glucose, Insulin, and C-Peptide Levels
40(1)
3.4.4 Assessment of Glucose Responsiveness Using GTT
40(1)
3.4.5 Survival Left-Side Nephrectomy and Tissue Removal
40(1)
3.4.6 Graft Histological and RNA Analyses
41(1)
3.5 Anticipated Results
42(2)
3.6 Discussion and Commentary
44(1)
3.7 Application Notes
45(1)
3.8 Summary Points
45(2)
References
46(1)
Chapter 4 Generation of Insulin-Producing Cells from Adult Mouse Intrahepatic Biliary Epithelial Cells In Vitro
47(18)
4.1 Introduction
48(1)
4.2 Experimental Design
49(1)
4.2.1 Generation of Insulin-Producing Cells from IHBECs
49(1)
4.3 Materials
49(4)
4.3.1 Reagents and Equipment
49(1)
4.3.2 Equipment
50(1)
4.3.3 Reagent Setup
51(1)
4.3.4 Adenovirus Transduction
52(1)
4.3.5 Reverse Transcription Polymerase Chain Reaction (RT-PCR)
52(1)
4.3.6 Immunocytochemistry
53(1)
4.3.7 Measurement of Insulin Production
53(1)
4.3.8 Analysis of Single Cells by RT-PCR
53(1)
4.4 Methods
53(6)
4.4.1 Step 1
53(1)
4.4.2 Mouse Biliary Epithelial Cell Isolation and Culture
53(2)
4.4.3 Step 2
55(1)
4.4.4 Step 3
55(2)
4.4.5 Step 4
57(1)
4.4.6 Step 5
57(1)
4.4.7 Evaluation
57(1)
4.4.8 Statistical Analysis
58(1)
4.5 Data Acquisition, Anticipated Results, and Interpretation
59(1)
4.6 Discussion and Commentary
59(3)
4.7 Summary Points
62(3)
Acknowledgments
62(1)
References
62(3)
Chapter 5 Technologies to Improve Human Islet Transplantation
65(12)
5.1 Introduction
66(1)
5.2 Materials
66(2)
5.2.1 Pancreas Procurement and Preservation
66(1)
5.2.2 Islet Isolation
66(1)
5.2.3 Islet Culture/Preservation (Optional)
67(1)
5.2.4 Islet Transplantation
68(1)
5.3 Methods
68(5)
5.3.1 Pancreas Procurement for Islet Isolation
68(1)
5.3.2 Pancreas Procurement from Heart-Beating Brain-Dead Donors
69(1)
5.3.3 Pancreas Procurement from Non-Heart-Beating Donors
70(1)
5.3.4 Pancreas Preservation
70(1)
5.3.5 Islet Isolation
71(1)
5.3.6 Islet Culture/Preservation (Optional)
72(1)
5.3.7 Islet Transplantation
72(1)
5.3.8 Potential Pitfalls and Troubleshooting
73(1)
5.4 Data Acquisition, Anticipated Results, and Interpretation
73(1)
5.4.1 Islet Isolations from Brain-Dead Donors
73(1)
5.4.2 Islet Isolations from Non-Heart-Beating Donors
74(1)
5.4.3 Islet Transplantation from Living Donors
74(1)
5.5 Discussion and Commentary
74(1)
5.6 Summary Points
75(2)
Acknowledgments
75(1)
References
75(2)
Chapter 6 Pancreatic Islet Isolation from Swine
77(24)
6.1 Introduction
78(1)
6.2 Materials
79(1)
6.2.1 Animals
79(1)
6.2.2 Younger Versus Older Pigs
79(1)
6.3 Equipment
80(1)
6.3.1 Major Equipment
80(1)
6.3.2 Small Equipment and Nondisposable Items
80(1)
6.3.3 Disposable Items
80(1)
6.4 Media
81(1)
6.5 Methods
81(15)
6.5.1 Harvesting the Pancreas
81(1)
6.5.2 Laboratory Setup
82(1)
6.5.3 Cleaning and Cannulating Pancreas
83(2)
6.5.4 Enzyme Preparation
85(4)
6.5.5 Polysucrose Purification of Tissue
89(3)
6.5.6 Counting Islet Equivalents
92(1)
6.5.7 Culture
93(1)
6.5.8 Assessment of Islet Quality
93(3)
6.6 Anticipated Results
96(1)
6.7 Discussion and Commentary
96(1)
6.8 Summary
97(4)
Acknowledgments
98(1)
References
98(3)
Chapter 7 Spinal Cord Injury Models and Cell Transplantation Therapy
101(20)
7.1 Introduction
102(1)
7.2 Experimental Design
102(1)
7.2.1 Pathophysiologic States of Spinal Cord Injury and Therapeutic Strategies
102(1)
7.2.2 Induction of Models Suitable for the Study Objectives
103(1)
7.2.3 Controls Required for the Experiments
103(1)
7.3 Materials and Methods
103(10)
7.3.1 Spinal Cord Section Model
103(3)
7.3.2 Spinal Contusion Injury Model
106(3)
7.3.3 Demyelination Model
109(2)
7.3.4 Cell Transplantation
111(2)
7.4 Data Acquisition, Anticipated Results, and Interpretation
113(4)
7.4.1 Function Assessment Methods
113(4)
7.5 Discussion and Commentary
117(1)
7.5.1 Ethical Aspects
118(1)
7.6 Application Notes
118(1)
7.7 Summary Points
118(3)
References
118(3)
Chapter 8 Cell Transplantation and Regeneration in Neurology
121(10)
8.1 Introduction
122(1)
8.2 Experimental Design
122(1)
8.3 Materials
123(1)
8.3.1 Preparation for Cell Suspension
123(1)
8.3.2 Stereotaxic Intracerebral Cell Transplantation (Rat)
124(1)
8.3.3 Encapsulated Cell Production
124(1)
8.4 Methods
124(3)
8.4.1 Preparation of Cell Suspension
124(1)
8.4.2 Stereotaxic Intracerebral Cell Transplantation (Rat)
125(2)
8.4.3 Making of Encapsulated Cells
127(1)
8.5 Data Acquisition, Anticipated Results, and Interpretation
127(1)
8.6 Discussion and Commentary
128(1)
8.7 Application Notes
128(1)
8.8 Summary Points
129(1)
8.9 Conclusions
130(1)
Acknowledgments
130(1)
References
130(1)
Chapter 9 Biocompatible Genetic Modification of Multipotent Mesenchymal Stromal Cells
131(24)
9.1 Introduction
132(3)
9.2 Materials
135(1)
9.2.1 Reagents
135(1)
9.2.2 Facilities/Equipment
136(1)
9.3 Methods
136(8)
9.3.1 Preparation of Pullulan-Spermine
136(2)
9.3.2 Preparation of Plasmid DNA
138(1)
9.3.3 Isolation and Culture of Primary Rat Bone Marrow MSCs
138(1)
9.3.4 Preparation of MSCs for Transfection in 24-Well Plates
139(1)
9.3.5 Pullulan-Spermine/DNA Anioplex Preparation and Anioplex Transfection
139(1)
9.3.6 WST-8 Assay
140(1)
9.3.7 Luciferase Assay
141(1)
9.3.8 Anioplex-Mediated Ex Vivo Transfection of MSCs Prior to Transplantation
141(1)
9.3.9 Lumbar Puncture Injection of MSCs
142(1)
9.3.10 Luciferase Assay
143(1)
9.4 Data Acquisition, Anticipated Results, and Interpretation
144(3)
9.5 Discussion and Commentary
147(3)
9.6 Application Notes
150(1)
9.7 Summary Points
150(5)
Acknowledgments
151(1)
References
151(4)
Chapter 10 Adipose Tissue as a Source of Mesenchymal Stem Cells for Hepatocyte Transplantation
155(18)
10.1 Introduction
156(1)
10.2 Materials
157(2)
10.3 Methods
159(5)
10.3.1 Isolation and Culture of AT-MSCs
159(1)
10.3.2 Flow Cytometry
159(1)
10.3.3 In Vitro Culture and Expansion of BM-MSC, BM-HSC, and HepG2 Cell Line
160(1)
10.3.4 RT-PCR
161(1)
10.3.5 Immunofluorescence
161(1)
10.3.6 Western Blot Analysis
161(1)
10.3.7 LDL Uptake
161(2)
10.3.8 Periodic Acid-Schiff Staining
163(1)
10.3.9 Ammonia Clearance
163(1)
10.3.10 Albumin Production
163(1)
10.3.11 CD105+ AT-MSC-Derived Hepatocyte Transplantation into Mice with CCl4-Induced Injury
163(1)
10.4 Anticipated Results
164(3)
10.4.1 Characterization of AT-MSCs
164(3)
10.5 Discussion and Commentary
167(2)
10.5.1 Hepatocyte Differentiation and Hepatic Regeneration
167(1)
10.5.2 Uncultured AT-MSCs
168(1)
10.6 Clinical Applications of Uncultured AT-MSCs
169(2)
10.6.1 Breast Reconstruction
169(1)
10.6.2 Cardiovascular Disease
170(1)
10.7 Summary Points
171(2)
Acknowledgments
171(1)
References
171(2)
Chapter 11 Culture Protocols for Producing Definitive Endoderm and Pancreatic Lineage Cells from Human ES or iPS Cells
173(10)
11.1 Introduction
174(1)
11.2 Experimental Design
175(2)
11.2.1 Materials
175(1)
11.2.2 Protocols
176(1)
11.3 Data Acquisition, Anticipated Results, and Interpretation
177(1)
11.4 Discussion and Commentary
178(1)
11.5 Application Notes
179(1)
11.6 Summary Points
180(3)
References
181(2)
Chapter 12 Human Placenta Cell Isolation
183(18)
12.1 Introduction
184(1)
12.2 Placenta Embryological Development
184(1)
12.3 Placenta Tissue Structure
185(1)
12.4 Placenta Immunological Properties
186(1)
12.5 Human Epithelial Cells from Amnion
186(1)
12.6 Human Mesenchymal Stromal Cells from Amnion
187(1)
12.7 Human Mesenchymal Stromal Cells from Chorion
188(1)
12.8 Materials
189(1)
12.9 Methods
190(4)
12.9.1 Preparation of Solutions for Isolation and Culture
191(1)
12.9.2 Preparation for Cell Isolation
191(1)
12.9.3 Release Amniotic Epithelial Cells
191(2)
12.9.4 Anticipated Results
193(1)
12.9.5 Release Amniotic Mesenchymal Cells
193(1)
12.9.6 Anticipated Results
194(1)
12.10 Placenta-Isolated Cell Characterization
194(1)
12.11 Cell Banking
195(1)
12.12 Discussion and Commentary
195(1)
12.13 Application Notes
196(1)
12.14 Summary Points
197(4)
Acknowledgments
198(1)
References
198(3)
Chapter 13 Liver Repopulation by Fetal Liver Stem/Progenitor Cells
201(18)
13.1 Introduction
202(2)
13.1.1 Background
202(1)
13.1.2 Cell Transplantation Strategies in Rodent Model Systems
202(1)
13.1.3 Fetal Liver Stem/Progenitor Cells as a Source for Liver Repopulation
203(1)
13.2 Experimental Design
204(1)
13.3 Animals
204(1)
13.4 Materials
204(2)
13.4.1 Cell Isolation
204(1)
13.4.2 Cell Transplantation
205(1)
13.4.3 Detection of Transplanted Cells
205(1)
13.5 Methods
206(6)
13.5.1 Cell Isolation
206(1)
13.5.2 Cell Transplantation
207(2)
13.5.3 Detection of Transplanted Cells
209(3)
13.6 Anticipated Results and Discussion
212(4)
13.6.1 Isolation of ED14 Fetal Liver Cells
212(1)
13.6.2 Transplantation of ED14 Fetal Liver Cells
212(1)
13.6.3 Detection of Transplanted DPPIV+ Cells
213(3)
13.7 Summary Points
216(3)
Acknowlegments
216(1)
References
216(3)
Chapter 14 Myoblast Transplantation: Techniques in Nonhuman Primates as a Bridge to Clinical Trials
219(18)
14.1 Introduction
220(1)
14.2 Experimental Design
221(1)
14.3 Materials
221(3)
14.3.1 Facilities/Equipment
221(1)
14.3.2 Reagents
222(1)
14.3.3 Cell Culture Material
222(1)
14.3.4 Surgical Material
223(1)
14.3.5 Cell Line
223(1)
14.4 Methods
224(5)
14.4.1 Cell Culture
224(3)
14.4.2 Cell Transplantation
227(2)
14.5 Data Acquisition, Anticipated Results, and Interpretation
229(2)
14.6 Discussion and Commentary
231(2)
14.7 Application Notes
233(1)
14.8 Summary Points
234(3)
References
234(3)
Acronyms and Abbreviations 237(4)
About the Editors 241(2)
Contributors 243(2)
Index 245
Alejando Soto-Gutierrez is an assistant professor in the Department of Surgery, Transplantation Section at the Children's Hospital of Pittsburgh. Nalu Navarro-Alvarez is a research scientist in the Columbia Center for Translational Immunology (CCTI) at Columbia University. Ira J. Fox is the director of the Center for Regenerative Therapies at Children's Hospital of Pittsburgh and the MacGowan Institute for Regenerative Medicine. He is also a professor of surgery and pediatric transplantation at the University of Pittsburgh School of Medicine.