PDF电子书：Stem Cells for Myocardial Regeneration Methods and Protocols 2010

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2010出版的 Methods in Molecular Biology 系列 vol.660
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2 ]9 J: E$ C: T+ L以下为该书前言：
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Preface  C6 |5 r* [1 o
The field of regenerative medicine is in its infancy state. Enthusiasm for the potential of0 i" O0 K6 I  B
organ regeneration lies with the potential pluripotency of stem cells to differentiate into
) \" `5 s# [5 z2 P$ y8 M$ Xvarious tissue types. This volume of Methods in Molecular Biology will focus on the use
9 t% ~* m3 A+ Z- t; L" F2 kof stem cells for myocardial repair and regeneration. The emphasis of this issue will be to8 b6 M0 D" @7 S: T/ w( a
provide basic scientists, translational investigators, and cardiologists a means to evaluate3 o6 G. c) b- F3 h+ E5 j
the efficacy and safety of stem cells in a standardized fashion for myocardial regeneration.: X! t( `; i2 j  {; \& z; z
Many different cell types have been considered for myocardial repair. Adult cardiomyocytes
; D! M$ q7 |  n% oare unable to survive even when transplanted into normal myocardium. The use5 I7 b' v- t3 d, y7 C" w8 O
of fetal or neonatal cardiomyocytes is not a feasible source of cells due to ethical concerns$ K/ g, ~; Z0 x. S6 E/ P$ ^
and donor availability. Therefore, the use of pluripotent stem cells has become the focus
6 g$ @. F8 ]$ {# t9 s2 E: V& Cof a cell source for myocardial repair and regeneration. A variety of stem cell types have
: j+ ^" P6 @4 v* Bbeen suggested to participate in myocardial repair. This has led the investigators to search; B  E/ d2 [  A" G! p
for the “optimal cell type for myocardial repair”. Reliable isolation of the cell source with& p/ R; {$ o9 f) z5 `; v# c( N
the ability to expand the cell population is a prerequisite. In the first section of this book,
, d. v6 G5 e' i4 A' H% n& mmethods for isolation of commonly used stem cells being investigated for myocardial
: i- ^# O3 k% n! H, Y, cregeneration are presented.+ R2 S: h' ]. _1 V. u3 N
Once a stem cell source has been selected, the stem cell needs to be tested in an appropriate
. I& O* d' Z) A/ P. h5 Fanimal model before being translated into clinical practice. Section 2 discusses both/ p. L9 P7 b9 ]. b! E
rodent and large animal models. The pros and cons of utilizing each of the models are; k; G( F0 a% l* V
discussed, as well as obtaining consistent myocardial pathology to test whether the stem
2 F7 W( s/ u% Q; c# ncells improve function. Techniques used to assess left ventricular function are described for& o  [& _" m* l/ L$ w; W6 h
both rodent and large animals, as well as methods to identify stem cells and their effect on& @: K1 r0 f. s" B0 h1 o
myocardial repair.
! P" q" e' z/ Y/ I$ d0 i9 cUnderstanding the developmental process of the human heart is paramount to developing
% \0 }5 O2 V7 D. Z- D5 i6 x  I( pstrategies for myocardial regeneration. Knowledge of the cellular components of$ a# u+ Y2 _: h8 ^+ N9 j# I
the heart and their response to injury is crucial in designing experiments and therapies for
& M8 F% c- s1 d3 imyocardial repair and regeneration. Discrepancies in results of stem cell differentiation
4 g4 q# B, _0 d& winto cardiomyocytes and its efficacy are commonly dependent on the interpretation of the$ d, M$ O  b: q
histological results. Section 3 reviews the histological characteristics of the developing and7 S4 h  P& J* b' q* n4 J
normal myocardium and provides the histological chronology of the heart following a! i2 d- Z+ D: N: [8 Z! ^$ }& m
myocardial infarction. Strategies for myocardial regeneration also include means to develop3 @9 Q6 ~! c$ p( z" ]& W+ c
a functional vascular system. It is important to discriminate between increases in capillary9 l3 E. Y" I; Z1 B
density that commonly do not increase blood flow and arteriogenesis that will lead to an9 L$ Y; @8 p+ _8 M. a- O$ Y
increase in blood flow. A detailed analysis of angiogenesis and methods to delineate the
- b( }0 V! u! Y7 h- u3 q, |types of vasculature produced by stem cells are also discussed in section 3.
) z; m+ v6 \, y; HOnce a stem cell is transplanted into the myocardium, it is of great importance to
: H3 {0 p. H# pdetermine its fate and to assure safety. MRI and molecular imaging enable the identification$ o  W: `- e6 h4 T+ `6 C; ^/ V
and tracking of transplanted stem cells. The use of superparamagnetic iron oxides to
# G+ L5 L- X6 Y$ ?label stem cells has enabled investigators to utilize MRI to assess the injection of stem cells0 |. S. P/ w" ~/ w8 f) B+ S
into the injured area and its effect on both segmental and global left ventricular function
. l0 }* x2 d, j, J2 }& \: o0 eand myocardial perfusion. Transfection of stem cells with a reporter gene allows the0 {# m1 l! z$ v3 q1 }# w
reporter probe to produce a signal detectable by commonly used imaging modalities.
2 m; u, D' G2 \' LMolecular genetic imaging is confined to viable stem cells and the population of stem cells& h  h# ?) w6 N+ {4 t
transfected, thus allowing for longitudinal tracking of stem cells. Molecular imaging has
  L1 }# r3 L+ V. Bbeen particularly useful in following embryonic stem cells and their propensity to form: u4 T0 f, O, l
teratomas. Recently, the beneficial effects of autologous stem cell therapy have been attributed
- h" d: W" }/ M/ g  Jto paracrine effects. The use of a genetic fate-mapping approach is reviewed in section
- a7 k. d/ i  t' E7 K' O4 to study adult cardiomyocyte replenishment following an injury. The use of the
( ^) D8 G3 A% P& Rtools in section 4 will allow investigators to address challenges of stem cell therapy such as- F, ]; B/ Y4 P1 S
stem cell retention, engraftment, and safety, and investigate the mechanisms of stem cell
. F8 ~- _: h2 a8 b- f9 u; utherapy.$ {* `/ R: y/ D  S/ u
The emphasis of myocardial regeneration has focused on improvements of left ventricular
/ Y! Q; {1 C3 A. S% d% C) V* yfunction; however, an electrically integrated transplanted stem cell with its surrounding; v4 ~$ x6 f# Z; f
environment is necessary to mitigate abnormal arrhythmias and optimize
- G, ~$ Q2 |& ]" t& a+ Q* ielectromechanical performance. Both in vitro assessment of cellular electrophysiological
& W3 x6 x9 s0 H) G$ x) C9 bproperties and cell-to-cell communication can be accomplished with multielectrode array# a% o6 E9 X0 H: d2 y! D
recordings and optical mapping. These studies can be complemented with either ex vivo) {! d. i9 i6 J% ]
optical mapping or in vivo electrophysiology studies. These methods are presented in$ G+ o: l& M" J1 A4 @. B' q0 S0 h1 M
section 5.7 V4 u/ T' p" i& F- R+ T3 I
Tissue engineering techniques have been used to enhance cell retention and create the
, R5 p  U6 J$ e* }- kmicroenvironment to allow for stem cell survival. More recently, the extracellular matrix. }1 Q4 M3 F! \& T; G
or functional groups derived from extracellular matrix proteins have been shown to influence  N9 p! C: l1 q7 g& L
stem cell binding, the production of growth factors by the stem cell, and stem cell% S* {0 p  t1 T: F1 o& t
differentiation. In the final section of this volume, a strategy for investigating the effects( M( y# L. g6 p& ]9 d
of the extracellular matrix on stem cell renewal and differentiation is presented., S! H! K' a1 F3 J) \: L
The methods presented in this volume of Methods in Molecular Biology attempt to
8 w) N$ g! n1 @* @1 ]highlight techniques and strategies to be utilized in investigating the many challenges that$ m2 _4 p  I% ]7 _$ ^" ^
need to be addressed before stem cell therapy can become a mainstream therapy for myocardial. a% R9 s* x- X8 ~  E: ?: i" a
regeneration.
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San Francisco, CA   b9 ]8 D& C2 j: g6 o- t& j1 _
Randall J. Lee
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