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

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& m  f# R8 [) K. h) K3 N; |7 M# C2010出版的 Methods in Molecular Biology 系列 vol.660
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以下为该书前言：) ]/ D5 t- z9 \8 G* m8 s9 G

% \" I: e/ r2 A0 E1 P) e8 `Preface
; Q4 |; n# p3 QThe field of regenerative medicine is in its infancy state. Enthusiasm for the potential of; F2 s2 s3 l: G- r
organ regeneration lies with the potential pluripotency of stem cells to differentiate into; U' ~0 a; T" Z
various tissue types. This volume of Methods in Molecular Biology will focus on the use( M1 J5 H8 [* s% O1 w6 g# {( N' Q
of stem cells for myocardial repair and regeneration. The emphasis of this issue will be to
! |4 V# t5 a# J! n- ?provide basic scientists, translational investigators, and cardiologists a means to evaluate
$ N2 E. f) Z8 M# D9 x2 M' q2 Ythe efficacy and safety of stem cells in a standardized fashion for myocardial regeneration.0 W% L6 t. M$ x0 o9 {. S
Many different cell types have been considered for myocardial repair. Adult cardiomyocytes
' w( h) p. l0 {9 e9 J5 X5 sare unable to survive even when transplanted into normal myocardium. The use. s7 x: L) Z- M+ l' h/ P+ R9 Q) f
of fetal or neonatal cardiomyocytes is not a feasible source of cells due to ethical concerns' C  J- j1 u6 o) g
and donor availability. Therefore, the use of pluripotent stem cells has become the focus( ^& Y9 e" o7 ?& i
of a cell source for myocardial repair and regeneration. A variety of stem cell types have
6 m! E$ }2 c, A# a! Dbeen suggested to participate in myocardial repair. This has led the investigators to search
5 s. V4 n/ S5 c2 O9 {for the “optimal cell type for myocardial repair”. Reliable isolation of the cell source with
6 W6 m9 t" T- ?+ u6 O1 m( Jthe ability to expand the cell population is a prerequisite. In the first section of this book,5 v/ n; s$ V) u  @' @
methods for isolation of commonly used stem cells being investigated for myocardial* {. q, `* L' C  v& Z1 W) S0 P
regeneration are presented.
# |9 H) a) {: u/ ^( S% I8 v9 {Once a stem cell source has been selected, the stem cell needs to be tested in an appropriate" W$ ?! J6 b/ g4 x& k
animal model before being translated into clinical practice. Section 2 discusses both; }4 p' O! V9 F( S. h
rodent and large animal models. The pros and cons of utilizing each of the models are) u7 r8 L8 `) H/ e  J; H5 A
discussed, as well as obtaining consistent myocardial pathology to test whether the stem
( P" g% n1 v8 }; S5 w& M* h- u) |0 Ocells improve function. Techniques used to assess left ventricular function are described for
- {0 F4 U2 Q" h% j; Sboth rodent and large animals, as well as methods to identify stem cells and their effect on4 z& `3 ~3 \# B' M7 t: i2 ~+ N( \  A
myocardial repair.
% v9 a7 @# C7 Q8 g9 I3 y, vUnderstanding the developmental process of the human heart is paramount to developing
5 f1 a" p3 q* H& s8 N% A5 O- K  \8 r' |strategies for myocardial regeneration. Knowledge of the cellular components of
2 A* m0 x% ^# C0 Y7 ]/ ~5 Sthe heart and their response to injury is crucial in designing experiments and therapies for
) t6 }0 I$ O' qmyocardial repair and regeneration. Discrepancies in results of stem cell differentiation" R6 d0 `4 O, |5 ]% `/ I
into cardiomyocytes and its efficacy are commonly dependent on the interpretation of the! K. q+ |# m* H% W( m% ^: e
histological results. Section 3 reviews the histological characteristics of the developing and- o5 i. |4 \$ X
normal myocardium and provides the histological chronology of the heart following a
/ y& d/ r/ v" ]+ `3 Y; y  Amyocardial infarction. Strategies for myocardial regeneration also include means to develop
& A3 o! R1 Z& i. V: \0 Wa functional vascular system. It is important to discriminate between increases in capillary8 j' p3 o: w5 m& ^# w3 ?; \
density that commonly do not increase blood flow and arteriogenesis that will lead to an0 s; p1 o! p1 G7 ~& V$ e- p3 j
increase in blood flow. A detailed analysis of angiogenesis and methods to delineate the
, N& g! F' H. u' xtypes of vasculature produced by stem cells are also discussed in section 3.' _5 f, H, O/ ]% {: _
Once a stem cell is transplanted into the myocardium, it is of great importance to! U1 m: R! M. P
determine its fate and to assure safety. MRI and molecular imaging enable the identification5 d% }- y( i# Z/ E
and tracking of transplanted stem cells. The use of superparamagnetic iron oxides to
2 c5 G' x; `2 elabel stem cells has enabled investigators to utilize MRI to assess the injection of stem cells6 G% A7 S4 p* E0 S/ O$ j
into the injured area and its effect on both segmental and global left ventricular function
/ Z0 m) M) w2 @/ F9 Z) A7 O0 sand myocardial perfusion. Transfection of stem cells with a reporter gene allows the# s( y* D& H6 Z9 d& H0 k
reporter probe to produce a signal detectable by commonly used imaging modalities.
% V4 a/ Z% K1 C6 \# q/ _7 ^% U4 SMolecular genetic imaging is confined to viable stem cells and the population of stem cells
+ Y- c; Z4 w7 w1 D  r; @5 h( g1 ntransfected, thus allowing for longitudinal tracking of stem cells. Molecular imaging has5 q( @# l/ }0 t0 J* L2 g
been particularly useful in following embryonic stem cells and their propensity to form  X! P2 b! Q+ G  n% g: H+ W% t7 j- ]
teratomas. Recently, the beneficial effects of autologous stem cell therapy have been attributed
$ c% N; N! o# l7 d  Y: ]2 Tto paracrine effects. The use of a genetic fate-mapping approach is reviewed in section
/ \! E% K+ ^% Z) T$ N4 to study adult cardiomyocyte replenishment following an injury. The use of the- A1 P' U  d& d$ z' a
tools in section 4 will allow investigators to address challenges of stem cell therapy such as4 v* y9 v- _& r+ X" A- Q. s; Q
stem cell retention, engraftment, and safety, and investigate the mechanisms of stem cell
; M2 h/ n# D( ]3 |6 Etherapy.
! d  i2 ?3 z) [3 ^# [The emphasis of myocardial regeneration has focused on improvements of left ventricular  P( I6 M8 n0 \; W. |
function; however, an electrically integrated transplanted stem cell with its surrounding, r7 T  a9 ~& I/ d+ r- p6 O$ }5 G& @
environment is necessary to mitigate abnormal arrhythmias and optimize) ~& t* e4 M& r/ r5 X" J
electromechanical performance. Both in vitro assessment of cellular electrophysiological! ?- V3 b9 v3 N
properties and cell-to-cell communication can be accomplished with multielectrode array
, M3 a9 W0 z* }' Urecordings and optical mapping. These studies can be complemented with either ex vivo9 k; h6 H5 t# t! C2 T/ c
optical mapping or in vivo electrophysiology studies. These methods are presented in
' f8 P6 m# N1 |' w$ Bsection 5.
" k4 b. x0 o% R3 KTissue engineering techniques have been used to enhance cell retention and create the
( \( n; i; X  cmicroenvironment to allow for stem cell survival. More recently, the extracellular matrix' G0 T; q4 F% c9 V: D
or functional groups derived from extracellular matrix proteins have been shown to influence
: Z; H) t# P2 }& y6 N8 x) Ostem cell binding, the production of growth factors by the stem cell, and stem cell
. Z$ x3 H8 z2 V. X2 U+ G3 _4 ldifferentiation. In the final section of this volume, a strategy for investigating the effects
8 G4 C5 g6 \- {9 l1 [: u/ K) Lof the extracellular matrix on stem cell renewal and differentiation is presented.$ G9 E- ^8 l7 E# T6 V( o' w
The methods presented in this volume of Methods in Molecular Biology attempt to  a2 L- N( x8 Y7 N) T
highlight techniques and strategies to be utilized in investigating the many challenges that# U9 A# N- [: |
need to be addressed before stem cell therapy can become a mainstream therapy for myocardial3 l0 s  s5 u1 D7 }1 u
regeneration.3 @! R4 s" B2 N: w' }2 x, C- C
( R8 h5 \3 M8 c4 ~7 Z/ X
San Francisco, CA
7 a/ g  K3 _3 _6 ?( w3 z5 pRandall J. Lee
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