Human Cell Culture. Primary Hematopoietic Cells

349 Pages·2002·3.558 MB·English

by  Koller M.R. (ed.), Palsson B.O. (ed.), Masters J.R. (ed.)

HUMANCELL CULTURE
VolumeIV:PrimaryHematopoieticCells
Human Cell Culture
Volume4
Human Cell Culture
Volume IV:
Primary Hematopoietic Cells
editedby
ManfredR.Koller
Oncosis,SanDiego, CA, U.S.A.
BernhardO.Palsson
University of California,
DepartmentofBioengineering, LaJolla, CA, U.S.A.
and
JohnR.W.Masters
University College London,
InstituteofUrology, London, U.K.
KLUWER ACADEMIC PUBLISHERS
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eBookISBN: 0-306-46886-7
Print ISBN: 0-792-35821-X
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Table ofContents
Chapter 1: Hematopoietic stem andprogenitorcells 1
Manfred R. Koller, Ph.D. Oncosis
Bernhard 0. Palsson, Ph.D. University of California -San Diego
Chapter2:In vitroT-lymphopoiesis 31
Michael Rosenzweig, Ph.D. New EnglandRegional Primate Center
David T. Scadden, M.D. Massachusetts General Hospital
Chapter 3: T-lymphocytes: Mature polyclonal and antigen-specific cell expansion 45
Bruce L. Levine, Ph.D. Naval Medical Research Institute
Katia Schlienger, M.D., Ph.D. Naval Medical Research Institute
Carl H. June, M.D. Naval Medical Research Institute
Chapter4: The culture, characterization, and triggering ofB lymphocytes 101
Gerrard Teoh, M.D. Dana-Farber Cancer Institute
Kenneth C. Anderson, M.D. Dana-Farber Cancer Institute
Chapter5: Monocytesandmacrophages 125
Ivan N. Rich, Ph.D. Richland Memorial Hospital
Chapter 6: Isolation and cultivation ofosteoclasts and osteoclast-like cells 147
Philip A. Osdoby, Ph.D. Washington University
Fred Anderson. Washington University
William Maloney, M.D. Washington University Medical Center
Patricia Collin-Osdoby,Ph.D. Washington University
Chapter 7: Isolationand culture ofhuman dendritic cells 171
Michael A. Morse, M.D. Duke University
H. Kim Lyerly, M.D. Duke University
Chapter 8: In vitro proliferation and differentiation ofCD34+ cells to neutrophils 193
James G. Bender, Ph.D. Nexell Therapeutics, Inc.
Chapter 9: Isolation and culture ofeosinophils 219
Helene F. Rosenberg, M.D. National Institutes of Health
Chapter 10: Isolationandculture ofmastcellsandbasophils 241
Peter Valent, M.D. University of Vienna
Chapter 11: Purification and culture oferythroid progenitor cells 259
Chun-Hua Dai, M.D. VA Medical Center
Amittha Wickrema, Ph.D. University of Illinois -Chicago
Sanford B. Krantz, M.D. Vanderbilt University Medical School
Chapter 12: In vitro development ofmegakaryocytes andplatelets 287
Marcus 0. Muench. Ph.D. Universityof California -San Francisco
Jae-Hung Shieh, Ph.D. New York Blood Center
Chapter 13: Perspectives.ethics, andclinical issuesin the useofprimary human cells 317
Mary Pat Moyer. Ph.D. InCelI, Inc.
Introduction
The daily production of hundreds of billions of blood cells through the process of
hematopoiesis is a remarkable feat of human physiology. Transport of oxygen to tissues,
blood clotting, antibody-and cellular-mediated immunity, bone remodeling, and a host of
other functions in the body are dependent on a properly functioning hematopoietic
system. As a consequence, many pathological conditions are attributable to blood cell
abnormalities, and a fair number of these are now clinically treatable as a direct result of
hematopoietic research.
Proliferation of hematopoietic stem cells, and their differentiation into the many
different lineages of functional mature cells, is highly regulated and responsive to many
environmental and physiological challenges. Our relatively advanced understanding of
this stem cell system provides potentially important insights into the regulation of
development in other tissues, many of which are now being acknowledged as stem cell-
based, perhaps even into adulthood. The recent public and scientific fanfare following
announcement of human embryonic stem cell studies suggests that stem cell research will
continue to be a relevant and exciting topic.
Recent advancements in primary human hematopoietic cell culture have led to
remarkable progress in the study of hematopoiesis, stem cell biology, immunology,
carcinogenesis, tissue engineering, and even in clinical practice for the treatment of
disease. This unique comprehensive volume in the Human Cell Culture Series
encompasses research methodology for the growth and differentiation of all types of
primary hematopoietic cells. Over the past decade, many new techniques have been
developed to propagate human cells for a number of hematopoietic lineages, utilizing
specific growth factors, stroma, medium additives, perfusion culture, and other strategies.
Each of twelve hematopoietic cell types is covered by a leading expert in the field,
providing insightful background information along with detailed current culture and
assay techniques. In addition to uses for research applications, current and future clinical
applications of large-scale culture methods are also discussed. Because the procurement
and processing of primary human tissues can pose a significant barrier to new researchers
in this field, this subject is covered in detail within each chapter. The final chapter is
intended to guide scientists through the significant regulatory and ethical implications
associated with use of human and fetal tissues. A consistent format with generous
inclusion of tables and figures enables readers to locate key information about each
cell/t issue type covered. Additionally, numerous literature citations provide a valuable
reference for students and professionals in the hematology, immunology, oncology, and
bioengineering fields. It is our goal to stimulate interest in the study of human
hematopoiesis, with the belief that new therapeutic solutions for a variety of diseases will
result.
Manfred R Koller
Chapter 1
Hematopoietic Stem and Progenitor Cells
1Manfred R Koller and 2Bernhard O Palsson
1Oncosis, 6199 Cornerstone Ct., Suite 111, San Diego, CA 92121 and2Department of
Bioengineering, UCSD, 9500 Gilman Dr, La Jolla, CA 92093-0412. Tel: [phone]. E-
mail: [email protected]
1. INTRODUCTION
The human body consumes a staggering 400 billion mature blood cells
every day, and this number increases dramatically under conditions of stress
such as infection or bleeding. A complex scheme of multilineage
proliferation and differentiation, termed hematopoiesis (Greek for blood-
forming), has evolved to meet this demand. This regulated production of
mature blood cells from primitive stem cells, which occurs mainly in the
bone marrow (BM) of adult mammals, has been the focus of considerable
research. Ex vivo models of human hematopoiesis now exist that have
significant scientific value and promise to have an impact on clinical
practice in the near future. This chapter introduces the reader to the
fundamental concepts of hematopoiesis, and provides information required
for the implementation of human stem and progenitor cell culture
techniques. The rest of this volume contains chapters which address the
isolation, culture, and utility of each of the major mature human
hematopoietic cell types.
1.1 Function and Organization of the Hematopoietic
System
There are approximately a dozen major types of mature blood cells
which are found in the body, depending upon the subdivision nomenclature
used (Fig. 1). These populations are divided into two major groups: the
1
2 KollerandPalsson
myeloid and lymphoid. The myeloid lineages include erythrocytes (red
blood cells), monocyte lineage-derived cells (eg. macrophages, osteoclasts,
and dendritic cells), the granulocytes (e.g. neutrophils, eosinophils,
basophils, and mast cells), and platelets (derived from non-circulating
megakaryocytes). Thymus-derived (T)-lymphocytes, BM-derived (B)-
lymphocytes, and natural killer (NK) cells constitute the lymphoid lineages.
Most mature blood cells exhibit a limited lifespan in vivo. Although some
lymphocytes are thought to survive for many years, it has been shown that
erythrocytes and neutrophils have lifespans of 120 days and 8 hours,
respectively [1]. As a result, hematopoiesis is a highly prolific process
which occurs throughout our lives to fulfill this demand.
Mature cells are continuously produced from progenitor cells which are
produced from earlier cells, which in turn originate from stem cells. At the
top (Fig. 1) are the very primitive totipotent stem cells, the majority of
which are in a nonproliferative state (G) [2]. These cells are very rare (1 in
0
105 BM cells), but collectively have enough proliferative capacity to last
several lifetimes [3,4]. Through some unknown mechanism(s), at any given
time a small number of these cells are actively proliferating, differentiating,
and self-renewing, thereby producing more mature progenitor cells while
maintaining the size of the stem cell pool. Whereas stem cells (by
definition) are not restricted to any lineage, their progenitor cell progeny do
have a restricted potential and are far greater in number. Therefore, as the
cells differentiate and travel from top to bottom in Fig. 1, they become more
numerous, lose self-renewal ability, lose proliferative potential, become
restricted to a single lineage, and finally become a mature functional cell of
a particular type.
1.2 Stem Cell Self-Renewal
Although stem cells have traditionally been thought to be capable of
unlimited self-renewal, new data suggest that this may not actually be the
case. For example, stem cells isolated from fetal liver, neonatal umbilical
cord blood (CB), and adult BM show a clear hierarchy of proliferative
potential [5]. One hypothesis that has been suggested is that the length of
telomeric DNA at the ends of chromosomes is shortened over time, acting as
a mitotic clock that triggers replicative senescence once telomeres reach a
threshold length [6]. In support of this hypothesis, longer telomeres and
greater telomerase activity (which extends telomeres) have been measured
in germline cells and tumor cells that do not exhibit replicative senescence
[7], Interestingly, telomerase activity is relatively high in stem cells,
although the activity does not appear to be great enough to impart
immortality [8]. While one study showed that introduction of telomerase
1. HematopoieticStemandProgenitorCells 3
Figure 1. The hematopoietic system hierarchy. It is believed that dividing pluripotent stem
cells may undergo self-r enewal to formdaughter stein cells without loss of potential (a matter
of current debate), or may experience a concomitant differentiation to form daughter cells
with more restricted potential. Continuous proliferation and differentiation along each lineage
results in the production of many ,mature cells. This process is under the control of many
growth factors (see Table I), and the sites of action for some of these are shown. The
mechanisms that determine which lineage a cell will develop into are not fully understood,
although many models have been proposed.