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

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8 U6 t, g% J9 l. _, [% G+ i9 y3 J. W2010出版的 Methods in Molecular Biology 系列 vol.660
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以下为该书前言：, x' P8 x+ P" o1 \5 j
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Preface9 b9 d( V: `# ]' C8 H3 d" [
The field of regenerative medicine is in its infancy state. Enthusiasm for the potential of' s* {7 p: Q/ G
organ regeneration lies with the potential pluripotency of stem cells to differentiate into' N: O. Y- M0 m/ ?/ Y1 w+ o
various tissue types. This volume of Methods in Molecular Biology will focus on the use
* ?2 m- N$ {1 D* w( s7 w* j: bof stem cells for myocardial repair and regeneration. The emphasis of this issue will be to+ F/ f1 G( q% V5 b$ b
provide basic scientists, translational investigators, and cardiologists a means to evaluate% o3 y/ ^/ }  D/ ^; \
the efficacy and safety of stem cells in a standardized fashion for myocardial regeneration.
) P/ M- H1 ~) O) qMany different cell types have been considered for myocardial repair. Adult cardiomyocytes( u! P2 O( P1 \* \% P
are unable to survive even when transplanted into normal myocardium. The use+ R$ c, J( F, M( Z! T: z' m6 p
of fetal or neonatal cardiomyocytes is not a feasible source of cells due to ethical concerns3 P! Y; R3 S1 F! ~% g
and donor availability. Therefore, the use of pluripotent stem cells has become the focus
* J9 }" D8 i9 ^; ?+ r; J9 L8 z: @of a cell source for myocardial repair and regeneration. A variety of stem cell types have
0 w5 h/ w2 J# l# Ubeen suggested to participate in myocardial repair. This has led the investigators to search
4 `$ l) _# [6 Afor the “optimal cell type for myocardial repair”. Reliable isolation of the cell source with
8 h( p- z+ d# x# B* ~the ability to expand the cell population is a prerequisite. In the first section of this book,
4 t" o( o/ E$ D; B* T+ O/ Tmethods for isolation of commonly used stem cells being investigated for myocardial
0 f6 `1 q# A+ _regeneration are presented.
3 S$ R3 H/ C  wOnce a stem cell source has been selected, the stem cell needs to be tested in an appropriate
* \/ q+ j4 p7 x2 zanimal model before being translated into clinical practice. Section 2 discusses both
$ v' i% }6 @9 x/ x9 u0 S  e8 crodent and large animal models. The pros and cons of utilizing each of the models are- e0 g- `" W  m. k( b  r* J
discussed, as well as obtaining consistent myocardial pathology to test whether the stem, R6 x! W; D! a, u1 s
cells improve function. Techniques used to assess left ventricular function are described for* K" s, W! Q$ T: y  V4 p# ~
both rodent and large animals, as well as methods to identify stem cells and their effect on
- A& ~+ C0 b4 O/ |8 omyocardial repair.: t$ v8 U8 o% W* P- R1 w* k
Understanding the developmental process of the human heart is paramount to developing" u1 h6 u7 L) B+ j% ]- Q
strategies for myocardial regeneration. Knowledge of the cellular components of# J- z' V9 p9 t9 L7 V( R
the heart and their response to injury is crucial in designing experiments and therapies for- r- B9 n% r0 S. b/ O9 g% Q
myocardial repair and regeneration. Discrepancies in results of stem cell differentiation& ?# M+ Q* a6 a6 G2 g: j1 p# b  _! \; e
into cardiomyocytes and its efficacy are commonly dependent on the interpretation of the  T9 O& q5 Q- s+ g6 z4 p
histological results. Section 3 reviews the histological characteristics of the developing and/ O4 J: a' w* O' j1 ~/ J
normal myocardium and provides the histological chronology of the heart following a( _; u. u5 z! o+ I  ~
myocardial infarction. Strategies for myocardial regeneration also include means to develop
& Z$ e8 m) {* d, Oa functional vascular system. It is important to discriminate between increases in capillary& E8 ?% ~$ K# v; G
density that commonly do not increase blood flow and arteriogenesis that will lead to an
3 |" M6 Q! m0 |+ w, o# Lincrease in blood flow. A detailed analysis of angiogenesis and methods to delineate the$ I, v2 q, |9 v9 c1 e( Z- K
types of vasculature produced by stem cells are also discussed in section 3.
; j: y" M* f5 j$ `" ]6 @Once a stem cell is transplanted into the myocardium, it is of great importance to7 S; Q8 F  n7 L/ b
determine its fate and to assure safety. MRI and molecular imaging enable the identification# m! U% X/ r, z/ s
and tracking of transplanted stem cells. The use of superparamagnetic iron oxides to$ f7 k" ^; z# C5 q! ~) P
label stem cells has enabled investigators to utilize MRI to assess the injection of stem cells: B( Z& e/ L) l$ |
into the injured area and its effect on both segmental and global left ventricular function; c# k# x  u2 ?6 o
and myocardial perfusion. Transfection of stem cells with a reporter gene allows the  d2 B, l, y9 T  P4 I
reporter probe to produce a signal detectable by commonly used imaging modalities., @) q" l# y6 ?9 a( z/ m3 w/ I
Molecular genetic imaging is confined to viable stem cells and the population of stem cells7 J% h. U/ V. ?  s
transfected, thus allowing for longitudinal tracking of stem cells. Molecular imaging has4 J1 d1 t3 }( _) u0 i% _+ Y1 [, p
been particularly useful in following embryonic stem cells and their propensity to form) c7 @- O! v4 g
teratomas. Recently, the beneficial effects of autologous stem cell therapy have been attributed
9 W3 y8 c/ d; n: a$ |0 N; C6 jto paracrine effects. The use of a genetic fate-mapping approach is reviewed in section+ \7 f) n1 D8 o7 w: L% n
4 to study adult cardiomyocyte replenishment following an injury. The use of the
7 q- E: X+ W, {1 E, ytools in section 4 will allow investigators to address challenges of stem cell therapy such as4 G2 R* p4 [! ?% x) d( p
stem cell retention, engraftment, and safety, and investigate the mechanisms of stem cell
" L4 H6 p% }! \$ ptherapy.8 c5 K$ K) C0 ?
The emphasis of myocardial regeneration has focused on improvements of left ventricular
. B. b* ^, o( T/ n1 Xfunction; however, an electrically integrated transplanted stem cell with its surrounding
. _- Z4 B; y4 y& p3 f8 Renvironment is necessary to mitigate abnormal arrhythmias and optimize
& b; K1 @! L1 ]! g! J3 {0 Helectromechanical performance. Both in vitro assessment of cellular electrophysiological3 b$ w- A. v- _! |% M- I
properties and cell-to-cell communication can be accomplished with multielectrode array8 B& }# `) K1 k6 Z2 m' y
recordings and optical mapping. These studies can be complemented with either ex vivo
& B# b/ z+ |; |6 A" C5 c) y& R4 ?optical mapping or in vivo electrophysiology studies. These methods are presented in: i: ^' M- L1 \5 o; R4 j
section 5.
) Z8 Z6 J& [+ W  n$ E3 rTissue engineering techniques have been used to enhance cell retention and create the$ |; N! @" ~0 H& E" ]  M  E8 B
microenvironment to allow for stem cell survival. More recently, the extracellular matrix: \! K% ~4 J0 c2 z% j
or functional groups derived from extracellular matrix proteins have been shown to influence
) J% F% M, b2 k- X5 L' hstem cell binding, the production of growth factors by the stem cell, and stem cell
6 X% {4 c& H3 g8 l9 _differentiation. In the final section of this volume, a strategy for investigating the effects  e8 e& S5 X) a3 W/ j1 B
of the extracellular matrix on stem cell renewal and differentiation is presented.
5 v1 O$ a* p* I9 `The methods presented in this volume of Methods in Molecular Biology attempt to
$ n0 ~( [* K, vhighlight techniques and strategies to be utilized in investigating the many challenges that
- ?  ]4 D* U: zneed to be addressed before stem cell therapy can become a mainstream therapy for myocardial
# h  s8 E' A' T% Nregeneration.% l' X9 _% U1 J" g- ~$ `

6 c4 g5 H' {( b) f8 OSan Francisco, CA
) W3 S2 l( C. v$ URandall J. Lee! c& v; t: B/ ~( Q' G* E$ n
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