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.6601 Q) S8 u: v6 K
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7 M0 U8 L; C5 {% W) a（请勿外传，以免引起不必要的版权纠纷）: I% L# z5 Y  D

# a- A. I% m) `* x. @( B! f以下为该书前言：# B5 `: L) v: Y2 q$ t, Q+ n

) V1 {" L  w; H) y/ x8 [Preface
/ o. `* G: q/ i! b$ Y1 g+ iThe field of regenerative medicine is in its infancy state. Enthusiasm for the potential of
4 R% _1 g# x- J# ]2 g& Horgan regeneration lies with the potential pluripotency of stem cells to differentiate into  r3 M0 Y3 K" q+ O' F0 F" s4 a
various tissue types. This volume of Methods in Molecular Biology will focus on the use- F! e! X0 v4 C  i$ c' P- Z
of stem cells for myocardial repair and regeneration. The emphasis of this issue will be to
# p- E4 _" f6 p: |; kprovide basic scientists, translational investigators, and cardiologists a means to evaluate
1 _( y9 E0 r( t, n% O6 Pthe efficacy and safety of stem cells in a standardized fashion for myocardial regeneration.
; R  h+ s* s" j( W  Y1 L/ GMany different cell types have been considered for myocardial repair. Adult cardiomyocytes
. U4 g4 t- |( f, [+ ]5 j9 u0 tare unable to survive even when transplanted into normal myocardium. The use
2 O7 n9 j5 w' f7 K% A  Z4 Rof fetal or neonatal cardiomyocytes is not a feasible source of cells due to ethical concerns
! D% V7 t1 E  R# g  Yand donor availability. Therefore, the use of pluripotent stem cells has become the focus* @) _6 b5 u  S
of a cell source for myocardial repair and regeneration. A variety of stem cell types have! o: d0 h; D0 W6 Y& e
been suggested to participate in myocardial repair. This has led the investigators to search2 d- W# c" e1 E4 @# X; J/ N1 k
for the “optimal cell type for myocardial repair”. Reliable isolation of the cell source with, c1 ~9 L: W; H( `
the ability to expand the cell population is a prerequisite. In the first section of this book,
) w; v$ E9 p0 `* m% Cmethods for isolation of commonly used stem cells being investigated for myocardial% s$ k! }7 d- K
regeneration are presented.
( ], G- s+ R: p3 q  n' xOnce a stem cell source has been selected, the stem cell needs to be tested in an appropriate  n8 x9 ^9 S0 \
animal model before being translated into clinical practice. Section 2 discusses both9 G% }2 K, O6 K2 H) d. O! M
rodent and large animal models. The pros and cons of utilizing each of the models are3 ?/ m% l3 v/ Q" K
discussed, as well as obtaining consistent myocardial pathology to test whether the stem. R: M4 l: N7 f' n: n
cells improve function. Techniques used to assess left ventricular function are described for1 M: I% X6 C7 Q+ s8 E3 `$ B6 I" i
both rodent and large animals, as well as methods to identify stem cells and their effect on
$ o4 {7 L. o+ ]% u( K# ]. C7 Nmyocardial repair.
! P; _1 `, A  T+ `1 l! e1 e6 a  ~Understanding the developmental process of the human heart is paramount to developing6 ~+ ^, }5 n% d, p# N" n3 _) O
strategies for myocardial regeneration. Knowledge of the cellular components of
8 R5 e- `: d% k2 Z; }: A$ G6 ?4 l* Qthe heart and their response to injury is crucial in designing experiments and therapies for
1 c) {/ }' A! J- z/ a1 V0 tmyocardial repair and regeneration. Discrepancies in results of stem cell differentiation
; b9 _  j0 X. t8 X& x2 L# pinto cardiomyocytes and its efficacy are commonly dependent on the interpretation of the1 h! r) r8 d" k8 {3 [' e
histological results. Section 3 reviews the histological characteristics of the developing and+ g6 }- A5 ^9 p2 [7 u* N* G
normal myocardium and provides the histological chronology of the heart following a/ [, y1 E1 y3 R  R/ C* Q
myocardial infarction. Strategies for myocardial regeneration also include means to develop" j) R: [! G1 j* Q7 ~
a functional vascular system. It is important to discriminate between increases in capillary' [: l0 F$ H  M8 A: y
density that commonly do not increase blood flow and arteriogenesis that will lead to an& @# T0 R0 o' p- g; |  S2 O
increase in blood flow. A detailed analysis of angiogenesis and methods to delineate the( ~% _, K! H7 H. c3 F; C
types of vasculature produced by stem cells are also discussed in section 3.
8 C% S9 J! H8 b: ^9 EOnce a stem cell is transplanted into the myocardium, it is of great importance to) `0 E( K7 {$ o3 O6 F
determine its fate and to assure safety. MRI and molecular imaging enable the identification# `7 E+ w3 m0 A& k
and tracking of transplanted stem cells. The use of superparamagnetic iron oxides to% j5 k0 P$ o  m; m
label stem cells has enabled investigators to utilize MRI to assess the injection of stem cells! R0 A" ?) `$ r& D
into the injured area and its effect on both segmental and global left ventricular function: M& ?- |5 x, O$ _4 t8 R
and myocardial perfusion. Transfection of stem cells with a reporter gene allows the
' M3 `/ G( y' ^4 N7 G& t4 Wreporter probe to produce a signal detectable by commonly used imaging modalities.
) A0 s/ a8 {2 D$ G7 o9 mMolecular genetic imaging is confined to viable stem cells and the population of stem cells
+ O4 ~" C4 |4 S5 Ztransfected, thus allowing for longitudinal tracking of stem cells. Molecular imaging has" D8 Z: I, a( K4 P; {
been particularly useful in following embryonic stem cells and their propensity to form" e/ K2 `0 G$ \/ I
teratomas. Recently, the beneficial effects of autologous stem cell therapy have been attributed1 A  m% `1 [& A% ~" e
to paracrine effects. The use of a genetic fate-mapping approach is reviewed in section
$ ~8 C1 ^% t4 S) t( d/ X4 to study adult cardiomyocyte replenishment following an injury. The use of the7 B* @3 T% S9 E* T
tools in section 4 will allow investigators to address challenges of stem cell therapy such as
9 \2 r9 ^) }% @; Zstem cell retention, engraftment, and safety, and investigate the mechanisms of stem cell( a" V, a3 T  \- }) o' c7 e' ~: O( Q
therapy.
0 E+ i! m) Y1 K" s4 n* GThe emphasis of myocardial regeneration has focused on improvements of left ventricular( G' g2 c0 b9 Z
function; however, an electrically integrated transplanted stem cell with its surrounding8 m6 h% F  Y& `6 A% z) I; u, o' f
environment is necessary to mitigate abnormal arrhythmias and optimize
( T2 R" N; j8 w) b, I5 Pelectromechanical performance. Both in vitro assessment of cellular electrophysiological( |) K4 [: F  y& w  z9 B
properties and cell-to-cell communication can be accomplished with multielectrode array: J2 v8 M5 r: S( _
recordings and optical mapping. These studies can be complemented with either ex vivo3 a: Y2 s$ l% ^7 y* Z( p& z
optical mapping or in vivo electrophysiology studies. These methods are presented in% i+ h7 P7 j6 [
section 5.
+ K: ]$ ~6 C/ p/ P& {7 u5 RTissue engineering techniques have been used to enhance cell retention and create the, H: j, Q5 }# j" a8 |* R( I/ J
microenvironment to allow for stem cell survival. More recently, the extracellular matrix) A$ ~0 \+ V% B3 g  p2 L! V3 w
or functional groups derived from extracellular matrix proteins have been shown to influence; F5 ]1 |! d, B/ t& z6 l5 H
stem cell binding, the production of growth factors by the stem cell, and stem cell! m$ ^" s$ ]& Z6 B3 r! ]# P
differentiation. In the final section of this volume, a strategy for investigating the effects3 t/ \; ~7 c- M+ u2 Z) Q
of the extracellular matrix on stem cell renewal and differentiation is presented.
. Y& a) o0 T2 c, Z- {/ O  b. UThe methods presented in this volume of Methods in Molecular Biology attempt to. v" K8 ~7 N4 L5 W& }9 L
highlight techniques and strategies to be utilized in investigating the many challenges that
4 i/ J( U, X, a4 R. pneed to be addressed before stem cell therapy can become a mainstream therapy for myocardial
% E' w( ~! Y  p: r( j) u6 B  yregeneration.
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San Francisco, CA
5 @; @" c+ c; r8 D/ L! _" ?Randall J. Lee
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