Cellular Therapy: Principles, Methods, and Regulations

  • ID: 2367112
  • October 2009
  • Region: Global
  • 675 Pages
  • AABB
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This manual was designed as a compendium of state-of-the-art practices and methods for cellular therapy to aid in the development and operation of a clinical cellular therapy facility. Designed for those in academia, government and industry, it is also an essential reference for those in regulatory affairs and quality assurance as well as for laboratory technologists, managers, directors, physicians and scientists. Through descriptions of the rationale and methodology for a variety of cell processing and evaluation techniques, it will assist laboratory staff in developing procedures that comply with applicable regulations and standards. In addition to numerous examples and templates for laboratory document preparation, several methods are provided that include a general overview of the critical steps, materials and equipment used in each process. Each section has been compiled and edited by a team of experts in the field, with subchapters contributed by specialists in the specific subject matter. Also included is a fully searchable CD-ROM with appendices, methods and forms in Microsoft Word format so they can be fully customized.

Chapters cover:
- Regulatory considerations:

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About the Editors

Regulatory Considerations for Manufacturing Cellular Therapies in the United States
Section Editors: Darin J Weber, PhD, and Victoria A Lake, BSc, BA, RAC
1 A Brief History of FDA Regulation of Human Cells and Tissues
Liana Harvath, PhD
HPCs from Placental/Umbilical Cord Blood and Peripheral Blood
Proposed Approach to Regulation of Cellular and Tissue-Based Products
Future Directions
2 The FDA Perspective on the Manufacturing, Production, and Processing of
Regulated Cellular Therapies
Sarah Kennett, PhD; Cynthia Porter, PhD; Eda Bloom, PhD; and Keith Wonnacott, PhD
General Tips
Current Good Manufacturing Practices
Manufacture of the Cellular Therapy Product
3 The Investigational New Drug Application
David H McKenna, Jr, MD; Victoria A Lake, BSc, BA, RAC; and Darin J Weber, PhD
Chemistry, Manufacturing, and Controls Section
Appendix 3-1 Sample Outline for Preparing the Chemistry, Manufacturing,
and Controls Section of an Investigational New Drug Application
4 Biologics License Applications for Cellular Therapies
Elizabeth Smith
Pre-BLA Submission Activities
Review Timelines
Appendix 4-1 Sample Outline for the Content and Organization of a Biologics License
5 Electronic Submissions of Cellular Therapy Investigational New Drug and Biologics
License Applications
Daniel P Offringa
Brief History
Advantages of Electronic Submissions over Paper
Initial Considerations
Building the Submission
Finalizing the Submission
6 Cellular Therapies Regulated as Combination Products
Joyce L Frey-Vasconcells, PhD
What Is the Definition of a Combination Product?
Who Determines Which Center Will Have Review Responsibility for a Combination
Can the Applicant Determine Which Center Will Review the Product?
What Regulations Need Clarification from the FDA?
7 Standards and Accreditation for Cellular Therapies
Carolyn A Keever-Taylor, PhD
Cellular Therapy Product Standards
Standards Harmonization
Achieving Accreditation
Quality Assurance for Cellular Therapy Facilities
Section Editors: Ellen Areman, MS, SBB(ASCP); Lizabeth Cardwell, MT(ASCP), MBA, RAC;
Karen Edward, BS, MT(ASCP); Cynthia Elliott, MT, HP(ASCP), CQA(ASQ); and
Michele W Sugrue, MS, MT(ASCP)SBB
Origins of Quality Concepts
Cellular Therapy Today
8 Quality and Process Control
Lizabeth Cardwell, MT(ASCP), MBA, RAC, and Michele W Sugrue, MS, MT(ASCP)SBB
Quality Control Process Model
Change Control
Process Verification and Validation
Process Simulation
Product Changeover (Line Clearance)
Product Segregation
Donor Eligibility
Recovery or Collection (See also Section V)
Tracking and Traceability
Storage Controls
Transportation Controls
Receipt, Predistribution Shipment, and Distribution of HCT/Ps
Appendix 8-1 Incoming Critical Supplies Log
Appendix 8-2 Sample Change-Control Form
Appendix 8-3 Document Format for General Process Validation Protocol
Appendix 8-4 Product Changeover Record
Appendix 8-5 Sample Product Tracking Form
Method 8-1 Maintaining a Corrective and Preventive Action Program
9 Human Resources in the Cellular Therapy Facility
Brenda Alder, MS, MT(ASCP)SBB; Lizabeth Cardwell, MT(ASCP), MBA, RAC;
Michele W Sugrue, MS, MT(ASCP)SBB; and Ellen Areman, MS, SBB(ASCP)
Personnel Requirements and Selection
Proficiency Testing
Continuing Education
Appendix 9-1 Sample Policy for Personnel, Training, and Competency
Appendix 9-2 Sample Job Description for Cellular Therapy Specialist
Appendix 9-3 Sample Job Description for Quality Assurance Specialist
Appendix 9-4 Sample Cellular Therapy Facility Training Checklist
Appendix 9-5 Sample Competency Assessment Policy
Appendix 9-6 Sample Cellular Therapy Facility Competency Assessment Checklist
Appendix 9-7 Sample Competency Assessment Form
Appendix 9-8 Sample Policy for a Continuing Education Program
10 Equipment Management in the Cellular Therapy Facility
Equipment Qualification
Equipment Quality Control
Records Review
Computer-Controlled Equipment
Appendix 10-1 Critical Equipment Selection Form
Appendix 10-2 Equipment Qualification Form
Appendix 10-3 New Equipment Qualification Checklist
Appendix 10-4 Equipment Activity Log
Appendix 10-5 Equipment Requalification Checklist
Method 10-1 Selecting and Qualifying Equipment
Method 10-2 Equipment Management
11 Quality Assurance of Supplies and Materials for Cellular Therapy Facilities
Marcia Swearingen, MS, MT(ASCP)SBB
Customer and Supplier Relations
Supplier Qualification
Receipt, Inspection, and Testing of Supplies
Appendix 11-1 Supplier Qualification Survey
12 Documents and Records
Betsy W Jett, MT(ASCP), CQA, CQM/OE(ASQ)
Documentation as a Value-Added Activity
Characteristics of an Effective Document
Document Management
Characteristics of an Effective Record
Record Management
13 Audits of Cellular Therapy Facilities
Sharon E Tindle, MS
External Audits
Internal Audits
Audit Schedule
Audit Management and Review
Auditor Qualification and Training
Auditing Methods
Auditing Resources
Method 13-1 Internal Audits for the Cellular Therapy Facility
14 Maintaining Safety in Cellular Therapy Facilities through Quality Assurance
Florinna Estioco Dekovic, MT(ASCP)BB, CQA(ASQ)
The Facility Designed for Security
Personal Hygiene Practices
Protection of Personnel in the Cellular Manufacturing Facility
Aseptic Technique
Emergency Preparedness and Action Plans
Contamination Control
Ergonomic Safety
Safety While Using Compressed Gas Cylinders
Material Safety Data Sheets and Hazardous Materials
Facilities: General Procedures
Section Editors: Robert A Preti, PhD, and John R Godshalk, MSE, MBA
15 Key Considerations and General Procedures for Facilities
Robert A Preti, PhD, and John R Godshalk, MSE, MBA
Manufacturing Area Requirements
Manufacturing Support Area Requirements
QC Laboratory and Microbiology Laboratory Requirements
Information Technology Infrastructure
Mechanical and Utility Requirements
Warehouse Requirements
16 Aseptic Processing
Angela Ondo, MT(ASCP)
The Environment
Validation of Aseptic Processing
Method 16-1 Aseptic Technique for Cell Processing
Method 16-2 Operating and Maintaining the Biological Safety Cabinet
17 Cross-Contamination Issues in the Manufacture of Cellular Therapy
John Finkbohner, PhD
Identifying Process Risks: Process Mapping
Minimizing Process Risks: Optimizing Manufacturing Unit Operations and Validating
Unit Operation Robustness
Minimizing Process Risks: Controlling Routine Operation Variables
18 Computer Systems in the Cellular Therapy Facility
Paula L Brown, MT(ASCP), JD
Computer System Selection
Computer Validation Plan
Appendix 18-1 Computer System Selection Plan
Appendix 18-2 Sample Computer System Installation Plan
19 Software Considerations for Cellular Therapy Products
John R Godshalk, MSE, MBA
Verification vs Validation
Software Validation Basics
Software Validation Performance
Risk Analysis
Regulations for Software Used in the Production of Cellular Therapy Products
20 Approaches to Validation for Cellular Therapy Products
Gary C du Moulin, PhD, MPH
Unique Requirements for the Validation of Cellular Therapy Products
Cellular Therapy Processes That Should Be Validated
Elements and Definition of Validation
Creating Procedures and Constructing a Protocol
Maintaining a State of Validation
Process or Product Changes
Process and Product Development for Cellular Therapeutics
Section Editors: Janice M Davis-Sproul, MAS, MT(ASCP)SBB, and Olive J Sturtevant,
21 Overview of Process Development
Jo Lynn Procter, MEd, MT(ASCP)SBB; Amy McDaniel, PhD; Diane Kadidlo, MT(ASCP)SBB;
Lizabeth Cardwell, MT(ASCP), MBA, RAC; Michele W Sugrue, MS, MT(ASCP)SBB;
and Douglas Padley, MT(ASCP)
Development Process Initiation
Development Team and Communication
Facility Assessment
Equipment Assessment
Ancillary Reagents and Materials Assessment
Process Assessment
Assay Development
Risk Assessment Methodology
Additional Points to Consider
Information Systems and Databases
Appendix 21-1 Example of a Development Study Report
Appendix 21-2 Template for a Design-Phase Transition Approval Form
Appendix 21-3 Example of a Preimplementation Review Checklist
Collection of Cells for Transplantation
Section Editors: Michele Cottler-Fox, MD; Edward Snyder, MD; and Zbigniew M Szczepiorkowski,
Collection of Cells for Transplantation
Background and Historical Perspective
Current State of the Field
Overview of Section V
Future Directions
22 Bone Marrow Collection
Thomas R Spitzer, MD
Development of Human Marrow Collection Technique
Collection Technique
Postoperative Care
Nucleated Cell Yield and Quality Control
Donor Complications
Additional Issues
Method 22-1 Harvesting Bone Marrow
23 Collection of Cellular Therapy Products by Apheresis
Michael L Linenberger, MD, FACP
HPC Mobilization Regimens and Kinetics
Instruments and Methodologies for HPC-A Collection
Standard Procedures for HPC-A Collections
Collecting HPC-A from Pediatric Donors
Adverse Effects and Complications of CD34+ Cell Mobilization and HPC-A Collection
Appendix 23-1 Examples of Treatment Plans from Three Institutions for Adult
Mononuclear Cell Collection by Leukocytapheresis
Appendix 23-2 Examples of Treatment Plans from Three Institutions for Adult Large-Volume
Mononuclear Cell Collection by Leukocytapheresis
Appendix 23-3 Example of a Treatment Plan for Modified Anticoagulation for Apheresis
Appendix 23-4 Example of an Apheresis Flow Sheet
Appendix 23-5 Cellular Therapy—Hematopoietic Progenitor Cell (HPC) Apheresis
Disclosure Form
Method 23-1 Mononuclear Cell Apheresis Collection Using COBE Spectra
24 Umbilical Cord Blood Collection
Julie G Allickson, PhD, MS, MT(ASCP)
Collection Methods Refined
Comparing Cord Blood Collection Methods
Microbial Contamination: In Utero vs Ex Utero
Other Factors That Affect UCB Collection Quality
Techniques to Improve Cord Blood Collection
Method 24-1 Collecting Cord Blood In Utero
Method 24-2 Collecting Cord Blood Ex Utero
25 Assessment of Collection Quality
Grace S Kao, MD
Donor Issues
Quality Assessment of Cellular Products Intended for Hematopoietic Progenitor Cell
Quality Assessment of Cellular Products Intended for Other Uses
Appendix 25-1 Examples of Quality Characteristics (ie, Acceptance Criteria)
of Cellular Therapy Products after Collection
Production: Basic Processing
Section Editors: Melissa Croskell, MT(ASCP); Kathy Loper, MHS, MT(ASCP); and
David H McKenna, Jr, MD
Product Handling
Collection Issues
Variables Affecting Product Manufacturing
Processing Endpoints
Blood Group Compatibility
Product Volume
Manipulation for Cryopreservation
26 Basic Cellular Therapy Manufacturing Procedures
Melissa Croskell, MT(ASCP); Kathy Loper, MHS, MT(ASCP); and David H McKenna, Jr, MD
Basic Procedures
Additional Platforms for Manipulations
Donor Lymphocytes for Infusion
Method 26-1 Reducing Plasma in Marrow and Hematopoietic Progenitor Cells
from Apheresis
Method 26-2 HES Sedimentation for Manual Red Cell Removal
Method 26-3 Volume Reduction of Peripheral Blood Progenitor Cells
for Cryopreservation
Method 26-4 Separating Bone Marrow Buffy Coat Using the COBE 2991
Method 26-5 Processing Marrow on Fenwal CS3000 Plus for Mononuclear
Cell Enrichment
Method 26-6 Processing Marrow Using the COBE Spectra for Mononuclear
Cell Enrichment
Method 26-7 Separating Marrow by the Density Gradient Method Using COBE 2991
27 Umbilical Cord Blood Processing
Philip H Coelho, BSME, and Kathy Loper, MHS, MT(ASCP)
Manual Processing of Cord Blood
Automated Processing of Cord Blood
Current State of the Field and Future Directions
Method 27-1 Automated Processing of Umbilical Cord Blood with
Biosafe Sepax System and Related Accessories
Method 27-2 Automated Volume Reduction of Umbilical Cord Blood Using the
AutoXpress System
Method 27-3 Processing Cord Blood with the Prepacyte System
28 Cryopreservation of Cellular Therapy Products
Allison Hubel, PhD
Elements of a Cryopreservation Protocol
Method 28-1 Cryopreserving Cellular Therapy Products in Freezing Bags
Method 28-2 Automated Controlled-Rate Freezing, Storage, and Retrieval of
Volume-Reduced Cord Blood Units Using the BioArchive System
29 Cryopreservation Containers for Cellular Therapy Products
Herbert M Cullis, MT, and Mary Dadone, PhD
Freezing Bags: General Considerations
Container Materials
Freezer Cassette Size
Freezer Temperature
Container Filling
Container Volume
Container Overwraps
Product Removal
30 Transportation and Shipping of Cellular Therapy Products
Donna Regan, MT(ASCP)
Product Procurement
Equipment and Cautions
Limitations and Recommendations
General Guidelines
Responsibilities of Personnel Involved with Transport
Quality Control: Dry Shippers
Paperwork and Labeling
Method 30-1 Shipping Cord Blood Units
31 Thawing and Infusing Cellular Therapy Products
Meghan Delaney, DO, and Richard L Haspel, MD, PhD
Process for Infusion of Thawed Cellular Therapy Products
Problems during Infusion
Prevention of Adverse Events from CT Product Infusion
Method 31-1 Thawing of Hematopoietic Progenitor Cells: Laboratory and Bedside
Method 31-2 Filtration of Cellular Therapy Apheresis Products
Method 31-3 Washing Cellular Therapy Products Using the COBE 2991
Method 31-4 Washing Cryopreserved Hematopoietic Progenitor Cells to Remove
Dimethyl Sulfoxide
Method 31-5 Reconstitutive Thawing: Preparing Frozen Cord Blood Products for
Method 31-6 Infusion of Hematopoietic Progenitor Cells in Adult Patients
Production: Complex or Extensive Processing
Section Editors: Melissa Croskell, MT(ASCP); Kathy Loper, MHS, MT(ASCP); and
David H McKenna, Jr, MD
32 Graft Modification: Cell Enrichment or Depletion
Safa Karandish, MT(ASCP)
Cell Isolation Methods
Cell Enrichment
Method 32-1 Selecting CD34+ Cells Using the Isolex 300i Magnetic Cell Selection System
Method 32-2 Selecting CD34+ Cells Using the CliniMACS System
33 Cell Separation by Counterflow Centrifugal Elutriation
Julie Edwards, BS, MT(ASCP)
Method 33-1 Counterflow Centrifugal Elutriation Using a Floor-Model Centrifuge
Method 33-2 Monocyte Enrichment Using the Elutra Cell Separation System Procedure
34 In-Vitro Cell Culture and Expansion
John D McMannis, PhD
HPC Expansion Overview
Umbilical Cord Blood Expansion
Culture Vessels and Devices
T-Cell Expansion and Immunotherapies
Method 34-1 Ex-Vivo Expansion of Selected Hematopoietic Progenitor Cells from
Cord Blood
35 T-Cell Expansion: Cytotoxic T Cells
Jerome Ritz, MD
Methods for In-Vitro Isolation and Expansion
36 Regulatory T Cells: Allograft Augmentation with Rapamycin-Generated Donor
CD4+ Th2 Cells
Daniel H Fowler, MD; Vicki Fellowes, MT(ASCP); and Hanh Khuu, MD
Current State of the Field
Future Directions
Method 36-1 General Overview of Current Th2 Cell Manufacturing:
Rapamycin-Generated Donor Th2 Cells
37 Natural Killer Cells
David H McKenna, Jr, MD, and Jeffrey S Miller, MD
Method 37-1 Production of Peripheral-Blood-Derived, CD3-Depleted,
IL-2-Activated Natural Killer Cells
38 Human Mesenchymal Stromal Cells
Karen Bieback, PhD, and Hermann Eichler, MD
Production of Human MSCs
Characterization of MSCs In Vitro
Genomic Stability
Clinical Potential
Method 38-1 Isolating and Characterizing Mesenchymal Stem or Stromal Cells
from Marrow
Method 38-2 Isolating and Characterizing Mesenchymal Stem or Stromal Cells
from Cord Blood
Method 38-3 Isolating and Characterizing Mesenchymal Stem or Stromal Cells from
Adipose Tissue
39 Generation and Characterization of Human Dendritic Cells for Therapy
Theresa L Whiteside, PhD
Summary of Procedure
Method 39-1 Generating and Characterizing Human Dendritic Cells for Cellular Therapy
40 Pancreatic Islet Cells
Elina Linetsky, MSc, MT; Aisha Khan, MS, MBA; and Camillo Ricordi, MD
Historical Perspective
Donor Selection
Islet Isolation
Islet Purification
Method 40-1 Isolating Human Islet Cells
Method 40-2 Preparing Pancreatic Digestion Media: SERVA Enzyme Preparation
Method 40-3 Preparing Pancreatic Digestion Enzyme: Collagenase and Thermolysin
41 Biorepositories
Kathy Loper, MHS, MT(ASCP), and Melissa Croskell, MT(ASCP)
Technical Points to Consider
Data Management
Ethical and Legal Issues
The Patient Perspective
Current Status
The Future
42 Regenerative Medicine Approaches for Cellular and Tissue Therapies
Bryan Tillman, MD, PhD; Jennifer L Olson, PhD; and James J Yoo, MD, PhD
Components of Regenerative Medicine
Regenerative Medicine Therapies
Assessment and Characterization of Cellular Therapy Products
Section Editors: Ellen Areman, MS, SBB(ASCP), and Lynn O'Donnell, PhD
Current State of the Field
Section Overview
Future Directions
43 Validation of Test Methods
Nina K Garlie, PhD
Performance Parameters
Appendix 43-1 Sample Validation Form
Method 43-1 Validating Test Methods
44 Cell Counts and Differentials of Cellular Therapy Products
Carolyn A Keever-Taylor, PhD
Marrow HPC Products
Apheresis HPC Products
Sampling and Determination of Product Volume
Instrument and Equipment Quality Control
Method 44-1 Partial Plasma Removal from Marrow Samples
45 Enumeration of CD34+ Cells by Flow Cytometry
D Robert Sutherland, MSc, and Michael Keeney, FIMLS, FCSMLS(D)
CD34+ Cell Enumeration Using Multiparameter Cytometry and Sequential Boolean Gating:
The ISHAGE Protocol
Critical Issues in CD34+ Cell Enumeration
Method 45-1 Flow Cytometric Enumeration of CD34+ Hematopoietic Progenitor Cells
According to ISHAGE Protocol
46 Pancreatic Islet Characterization
Elina Linetsky, MSc, MT
Method 46-1 Enumerating Pancreatic Islets by Dithizone Staining
Method 46-2 Determining Pancreatic Islet Function by Insulin Release
47 Assessment of Viability and Apoptosis in Cellular Therapy Products
Nicholas Greco, PhD, and Lynn O'Donnell, PhD
Method 47-1 Determining Cellular Viability Using Trypan Blue
Method 47-2 Determining Viability by Fluorescence Microscopy Using Acridine
Orange and Propidium Iodide
Method 47-3 Assessing Viability by Measuring Apoptotic Cells Using Flow Cytometry
48 Colony-Forming Cell Assays for Determining Potency of Cellular Therapy Products
Emer Clarke, PhD
Method 48-1 Hematopoietic Colony-Forming Cell Assays
49 Bioluminescence Assays for Assessing Potency of Cellular Therapy Products
Karen M Hall, MT(ASCP), and Ivan N Rich, PhD
Calibration and Standardization of the Potency Assay
Cell Populations
Method 49-1 Measuring Proliferation Using Bioluminescence-Based ATP Assays
50 Assessing the Function of Cellular Therapy Products by Measurement of
Cytoplasmic Aldehyde Dehydrogenase
N Rebecca Haley, MD
Characteristics of Hematopoietic Cells with High ALDH
Method 50-1 Analyzing ALDH-Bright Cells by Flow Cytometry
51 Performance of Cytotoxicity Assays for Cellular Therapy Products
Ellen Areman, MS, SBB(ASCP)
Method 51-1 Assessing Cytotoxic T- or Natural Killer Cell Function: 51Chromium
Release Assay
Method 51-2 Assessing Cytotoxic T- or Natural Killer Cell Function:
CD107A Degranulation Assay
52 Assessing Cellular Therapy Products for Microbial Contamination
Hanh Khuu, MD
Special Challenges Pertaining to Sterility Testing of CT Products
Interpretation of Results
Future Directions
Method 52-1 Cellular Therapy Product Sterility Testing (As Described in 21 CFR 610 12)
Method 52-2 Cellular Therapy Product Sterility Testing Using an Automated
Microbial Detection System (Alternate Method)
53 Detection of Mycoplasma Contamination
Peter Bugert, PhD
Common Characteristics of Mycoplasma Species
Methods for Mycoplasma Detection
Validation of New Methods for Mycoplasma Detection
Future Directions of PCR-Based Mycoplasma Detection
Method 53-1 Determining Color-Changing Units for Microbiologic
Culture-Based Mycoplasma Detection
Method 53-2 Detecting Mycoplasma by Conventional PCR
54 Endotoxin Testing of Cellular Therapy Products
Diane Kadidlo, MT(ASCP)SBB
Method 54-1 Kinetic-Chromogenic LAL Assay
55 The Use of Blood Grouping for Identity Testing of Cellular Therapy Products
Ellen Areman, MS, SBB(ASCP)
56 HLA Typing for Cellular Product Characterization and Identity Testing
Terry O Harville, MD, PhD
HLA Terminology
Serologic, “Low-Resolution” HLA Typing
HLA Typing by DNA Analysis Techniques
Medium-to-High-Resolution HLA Typing by SSO
High-Resolution HLA Typing by SSP PCR Technique
HLA Typing by Sequence-Based Technology
Comparisons of DNA Typing Techniques
HLA Typing as an Identity Test
Appendix 56-1 Selected Internet Resources on the HLA System and HLA Typing
Method 56-1 Serologic HLA Typing by Complement-Dependent Cytotoxicity
Method 56-2 Performing Medium-to-High-Resolution HLA DNA Typing by SSO Luminex
Method 56-3 High-Resolution HLA Typing by SSP-PCR
Appendix 1 Global Cellular Therapy Regulatory Bodies and Organizations
Appendix 2 Glossary

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