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
  G9 T. j8 H: M3 @与本站朋友分享。
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4 U6 E5 \- ]- X以下为该书前言：
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Preface
& _2 H2 \$ Y% f" w2 I" xThe field of regenerative medicine is in its infancy state. Enthusiasm for the potential of
$ A) b/ U# s% iorgan regeneration lies with the potential pluripotency of stem cells to differentiate into
+ U- a2 t) i+ t0 Kvarious tissue types. This volume of Methods in Molecular Biology will focus on the use
0 W  `  F8 V9 A+ }0 l+ |% V0 r. }of stem cells for myocardial repair and regeneration. The emphasis of this issue will be to' m$ Q2 O0 G- S0 b2 z4 f. W. g- q
provide basic scientists, translational investigators, and cardiologists a means to evaluate
! v7 x. W7 t8 s" V' h7 [the efficacy and safety of stem cells in a standardized fashion for myocardial regeneration.
" w; @! n/ k6 g! wMany different cell types have been considered for myocardial repair. Adult cardiomyocytes
& P& w4 o& F# iare unable to survive even when transplanted into normal myocardium. The use4 V& W. D0 x7 n3 z" P  a7 X) v
of fetal or neonatal cardiomyocytes is not a feasible source of cells due to ethical concerns
6 J' ?$ h* T* V# l' B' ?% Mand donor availability. Therefore, the use of pluripotent stem cells has become the focus
3 e/ v% g. J, U" B0 D* `9 _of a cell source for myocardial repair and regeneration. A variety of stem cell types have
3 E/ @/ }( |. t  d4 }' G( dbeen suggested to participate in myocardial repair. This has led the investigators to search
( `- E" ?4 ]. M" C) Afor the “optimal cell type for myocardial repair”. Reliable isolation of the cell source with
; m; m) U& Q! Q3 {# N; d# `' \the ability to expand the cell population is a prerequisite. In the first section of this book,
1 o7 \- z! }0 pmethods for isolation of commonly used stem cells being investigated for myocardial
) n) _4 W. Y, M* o$ P2 Pregeneration are presented.
8 t8 l: E4 x8 g  ROnce a stem cell source has been selected, the stem cell needs to be tested in an appropriate
- u/ |2 A% H* {  U# Z* Uanimal model before being translated into clinical practice. Section 2 discusses both  T" w9 [% _; c9 X
rodent and large animal models. The pros and cons of utilizing each of the models are
: B1 t8 }' u* y* H, Ldiscussed, as well as obtaining consistent myocardial pathology to test whether the stem
( r9 ~# x$ o. v; Ecells improve function. Techniques used to assess left ventricular function are described for
2 C! |! Q+ l. ]7 P+ y/ xboth rodent and large animals, as well as methods to identify stem cells and their effect on
) p# y( j& x2 w+ j9 U( R' y* F' _; Cmyocardial repair.& N, S* O& ~1 }9 K2 z
Understanding the developmental process of the human heart is paramount to developing2 T4 G6 n- Y/ C
strategies for myocardial regeneration. Knowledge of the cellular components of
- i6 x9 s. r1 P7 kthe heart and their response to injury is crucial in designing experiments and therapies for& q  p8 D8 @  G
myocardial repair and regeneration. Discrepancies in results of stem cell differentiation
1 }% w2 N) I9 m) d$ sinto cardiomyocytes and its efficacy are commonly dependent on the interpretation of the3 Y# d2 }+ r& V1 m% J
histological results. Section 3 reviews the histological characteristics of the developing and
" b8 u; s8 r# l  B4 y' snormal myocardium and provides the histological chronology of the heart following a
7 Z% j4 P* Q3 O8 c7 ?myocardial infarction. Strategies for myocardial regeneration also include means to develop; C9 ~7 t$ {) p4 M5 Z4 m
a functional vascular system. It is important to discriminate between increases in capillary. h9 ^0 B$ x3 [; K7 N# k* u" v. A
density that commonly do not increase blood flow and arteriogenesis that will lead to an
# {# o9 l$ C! M$ b1 |# l- J6 Nincrease in blood flow. A detailed analysis of angiogenesis and methods to delineate the) A$ y- U% [' s. A4 y3 Y
types of vasculature produced by stem cells are also discussed in section 3.8 ~9 @) f, o4 g( \4 ~
Once a stem cell is transplanted into the myocardium, it is of great importance to/ p0 S5 X( e0 b0 }0 |) G2 d0 `3 j
determine its fate and to assure safety. MRI and molecular imaging enable the identification
" J& Y( x& S+ R' M" Xand tracking of transplanted stem cells. The use of superparamagnetic iron oxides to3 ]/ n7 @+ z) j! t# b4 e2 f% f
label stem cells has enabled investigators to utilize MRI to assess the injection of stem cells
/ w$ ?# h9 ?% n1 A: T, ^2 f6 g2 dinto the injured area and its effect on both segmental and global left ventricular function
8 J) u, w7 \: U* O9 X$ f7 jand myocardial perfusion. Transfection of stem cells with a reporter gene allows the& z" F5 r: \8 y1 A+ C( Q
reporter probe to produce a signal detectable by commonly used imaging modalities.
  B& V3 y3 Y/ A6 {( a% g# E0 MMolecular genetic imaging is confined to viable stem cells and the population of stem cells4 Q5 a/ d8 L; N* @9 i
transfected, thus allowing for longitudinal tracking of stem cells. Molecular imaging has
7 ~6 T! ?# [& v) ~9 ?! u; zbeen particularly useful in following embryonic stem cells and their propensity to form
# x5 M$ C4 U3 d6 @* J! eteratomas. Recently, the beneficial effects of autologous stem cell therapy have been attributed2 w, i3 C7 d9 P3 `8 m; m8 H
to paracrine effects. The use of a genetic fate-mapping approach is reviewed in section
1 V- ]; d9 m/ O* `9 {" I) ]4 to study adult cardiomyocyte replenishment following an injury. The use of the0 L2 j/ `1 {! y5 l
tools in section 4 will allow investigators to address challenges of stem cell therapy such as
4 Y5 I/ S! n' P9 U: N( D7 ystem cell retention, engraftment, and safety, and investigate the mechanisms of stem cell
- G, R: {1 E( I6 g" q1 c9 N( d( F  Stherapy.
1 B# K! s* u1 @' ]6 }The emphasis of myocardial regeneration has focused on improvements of left ventricular2 k* O; e5 b* ?$ [3 c- V
function; however, an electrically integrated transplanted stem cell with its surrounding
' M! A. ]* n8 P" |environment is necessary to mitigate abnormal arrhythmias and optimize
4 |! W3 F2 A! @, b* u/ W/ ~electromechanical performance. Both in vitro assessment of cellular electrophysiological
1 x) [9 {7 M' ?properties and cell-to-cell communication can be accomplished with multielectrode array
, C" }" l  z2 l, i2 a+ _* Hrecordings and optical mapping. These studies can be complemented with either ex vivo  f  m- {! I2 ?% ?, Y
optical mapping or in vivo electrophysiology studies. These methods are presented in' l( ?  c& v# o/ f/ S" d
section 5.; J  g) u; I' ^2 ]7 P1 q- n# U
Tissue engineering techniques have been used to enhance cell retention and create the
3 a4 d7 G9 a( A: Q5 D- Gmicroenvironment to allow for stem cell survival. More recently, the extracellular matrix
1 B2 _. e& F! A+ Q' ]( L6 L0 r6 Gor functional groups derived from extracellular matrix proteins have been shown to influence, n' _4 p  e# A6 P4 Z; M+ A0 v5 S
stem cell binding, the production of growth factors by the stem cell, and stem cell0 K7 p6 O7 N  k) c4 [0 m
differentiation. In the final section of this volume, a strategy for investigating the effects  G$ S" Y) A" k( A3 V% T3 v% V) s
of the extracellular matrix on stem cell renewal and differentiation is presented.4 `$ }9 i( L1 }' f$ Y
The methods presented in this volume of Methods in Molecular Biology attempt to
; g/ Q2 b' m; J5 Z, }7 m! ahighlight techniques and strategies to be utilized in investigating the many challenges that' r6 `1 X9 u8 _. L' h
need to be addressed before stem cell therapy can become a mainstream therapy for myocardial& ~; [( n: \- \' v
regeneration.
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0 \0 G, K$ ^; l9 P! MSan Francisco, CA
' l  v% G: \7 s( M! l( P) I/ ~Randall J. Lee
7 J6 W7 O5 `6 I* ]1 C# B+ I' T+ N" \1 U

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