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

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, v8 X5 J" }: i2010出版的 Methods in Molecular Biology 系列 vol.660
8 e% z) e" A) i0 w  K* f与本站朋友分享。
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& v: Z. D+ I+ Z以下为该书前言：: X; Q3 H3 B* a) K( ~( y- B

0 o4 r) d- C& \# a  j8 T7 kPreface
8 H; ?6 O: H$ U/ [- {* FThe field of regenerative medicine is in its infancy state. Enthusiasm for the potential of- W; ^- |/ a4 J. E, m$ g5 Z; b* p/ H
organ regeneration lies with the potential pluripotency of stem cells to differentiate into5 z& x6 M+ }' ~/ Q: I: x  o1 N3 I4 A
various tissue types. This volume of Methods in Molecular Biology will focus on the use
" k$ I8 A2 p/ t" B1 F2 Qof stem cells for myocardial repair and regeneration. The emphasis of this issue will be to+ o3 @" u! [1 j1 t: J3 ?1 u2 Z9 y3 B
provide basic scientists, translational investigators, and cardiologists a means to evaluate
0 i: L! F' y- w# d7 Z: Ithe efficacy and safety of stem cells in a standardized fashion for myocardial regeneration.
" Z  G6 @& G  m: N7 mMany different cell types have been considered for myocardial repair. Adult cardiomyocytes
6 I1 L  v; z) Gare unable to survive even when transplanted into normal myocardium. The use
+ u  y/ I" e$ d4 Tof fetal or neonatal cardiomyocytes is not a feasible source of cells due to ethical concerns8 @: K: t  H) s$ X  N
and donor availability. Therefore, the use of pluripotent stem cells has become the focus
) F2 \: ~* P; e3 ~% \0 c6 Kof a cell source for myocardial repair and regeneration. A variety of stem cell types have8 W; i9 L: b2 @2 |
been suggested to participate in myocardial repair. This has led the investigators to search
& X, e( H' {4 Cfor the “optimal cell type for myocardial repair”. Reliable isolation of the cell source with
5 T, j; {+ i; ^: f/ k1 tthe ability to expand the cell population is a prerequisite. In the first section of this book,
3 N4 a3 C$ N( A/ E, v: L/ Gmethods for isolation of commonly used stem cells being investigated for myocardial
8 B1 m& v/ o5 e  s7 [. iregeneration are presented.
, [5 g$ `7 _3 A9 uOnce a stem cell source has been selected, the stem cell needs to be tested in an appropriate5 w' B2 r# L: o5 V7 A4 H
animal model before being translated into clinical practice. Section 2 discusses both
' y# a! ^3 k8 ]+ grodent and large animal models. The pros and cons of utilizing each of the models are
+ \  h; ]; q' }9 ^% x5 s; K4 b3 udiscussed, as well as obtaining consistent myocardial pathology to test whether the stem
9 h" ~8 ~2 m& T2 tcells improve function. Techniques used to assess left ventricular function are described for) s& h( R) i* J1 e9 N/ C
both rodent and large animals, as well as methods to identify stem cells and their effect on8 p9 V/ @6 d6 R* @( e% L" }
myocardial repair.
9 F3 p6 |: V& o, S  K; O2 P5 fUnderstanding the developmental process of the human heart is paramount to developing
. {! q) i" b  i! ]% p- v1 tstrategies for myocardial regeneration. Knowledge of the cellular components of
3 R2 g* f) \1 k9 \) O# Pthe heart and their response to injury is crucial in designing experiments and therapies for
2 `2 x; U: R  ^  [. mmyocardial repair and regeneration. Discrepancies in results of stem cell differentiation& p9 `/ m% }5 ^3 O# }! R" d: g7 V! L9 J
into cardiomyocytes and its efficacy are commonly dependent on the interpretation of the' v& g% a! \+ e/ x& J
histological results. Section 3 reviews the histological characteristics of the developing and) h9 e$ v! T: m/ ?5 S  F
normal myocardium and provides the histological chronology of the heart following a) _: h: q3 O+ t5 c4 O+ t8 S
myocardial infarction. Strategies for myocardial regeneration also include means to develop2 t" H! a$ u1 c+ b! @; p
a functional vascular system. It is important to discriminate between increases in capillary8 s9 z" x. ]# t4 N. A
density that commonly do not increase blood flow and arteriogenesis that will lead to an  Z9 x- u- X6 p+ h( R& `4 i! D% H
increase in blood flow. A detailed analysis of angiogenesis and methods to delineate the
6 Y6 d* c* V7 Utypes of vasculature produced by stem cells are also discussed in section 3.4 g2 r6 M; ~. J0 h
Once a stem cell is transplanted into the myocardium, it is of great importance to
( v! J; @9 Q1 a! b0 a# b8 Edetermine its fate and to assure safety. MRI and molecular imaging enable the identification
+ X- ~8 L: \: d$ P$ k% p( Band tracking of transplanted stem cells. The use of superparamagnetic iron oxides to) ^& L! [) x6 L
label stem cells has enabled investigators to utilize MRI to assess the injection of stem cells1 u, J, m6 I+ h- z8 |: [
into the injured area and its effect on both segmental and global left ventricular function( A" j! l( A, b9 e( G! ~3 j
and myocardial perfusion. Transfection of stem cells with a reporter gene allows the
! M; q* A' B/ l# sreporter probe to produce a signal detectable by commonly used imaging modalities.
9 I6 k7 @( x/ }& I; AMolecular genetic imaging is confined to viable stem cells and the population of stem cells
. @' L6 z& l6 z/ d! v2 Xtransfected, thus allowing for longitudinal tracking of stem cells. Molecular imaging has
" t  P* e& o/ `# X: F2 jbeen particularly useful in following embryonic stem cells and their propensity to form
/ P" u7 B: y$ u& }teratomas. Recently, the beneficial effects of autologous stem cell therapy have been attributed
7 x2 M2 b9 @5 K0 ]4 k! S  Vto paracrine effects. The use of a genetic fate-mapping approach is reviewed in section
! m7 n4 X7 d+ k; j1 G4 to study adult cardiomyocyte replenishment following an injury. The use of the! c5 v) b1 v) `% I! Z
tools in section 4 will allow investigators to address challenges of stem cell therapy such as
: T5 u1 \+ k! ~/ istem cell retention, engraftment, and safety, and investigate the mechanisms of stem cell
: M& _0 F: X, o; ]7 L. Gtherapy.
- y4 [" p- R' ^( Q& R7 Q: F% OThe emphasis of myocardial regeneration has focused on improvements of left ventricular1 X: v3 A1 L8 G; G
function; however, an electrically integrated transplanted stem cell with its surrounding
5 w6 ]: [$ q1 Z0 l+ D- C, l* r* E$ nenvironment is necessary to mitigate abnormal arrhythmias and optimize7 c5 Q. ~& H6 w7 m
electromechanical performance. Both in vitro assessment of cellular electrophysiological
4 r$ Q: M9 }. E; ^' Rproperties and cell-to-cell communication can be accomplished with multielectrode array' B& C, A0 [; {% _
recordings and optical mapping. These studies can be complemented with either ex vivo2 s$ s* q. R0 |4 l9 A1 [- C1 e& f9 b
optical mapping or in vivo electrophysiology studies. These methods are presented in3 ?+ I' t+ k4 o7 t% [" ~
section 5.
6 p/ T4 W, ~; mTissue engineering techniques have been used to enhance cell retention and create the
1 \" X3 u/ a2 t, x# ?! |microenvironment to allow for stem cell survival. More recently, the extracellular matrix9 N% k' n2 o0 m% }) a+ n1 e
or functional groups derived from extracellular matrix proteins have been shown to influence0 I9 m2 E/ ]4 \! U/ ~  Z
stem cell binding, the production of growth factors by the stem cell, and stem cell( G" D) J0 e# ^- l8 s/ P( A
differentiation. In the final section of this volume, a strategy for investigating the effects* [0 o6 h* G# s3 t, B
of the extracellular matrix on stem cell renewal and differentiation is presented.
, ^* G6 E8 y; a5 @4 R* zThe methods presented in this volume of Methods in Molecular Biology attempt to
5 w/ {1 \0 T. I9 G1 `$ d. Hhighlight techniques and strategies to be utilized in investigating the many challenges that
! n6 o) \' F# g( f6 sneed to be addressed before stem cell therapy can become a mainstream therapy for myocardial
% L$ Z2 d' {4 o4 s* f% |" Oregeneration.0 Q- Y# I3 y; \1 Y8 h

9 ~* |) a  t( M5 d+ bSan Francisco, CA 7 j2 e0 Y& V: z8 ~; ?1 V
Randall J. Lee4 V% x9 x% }" P
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