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本帖最后由 细胞海洋 于 2009-8-13 12:01 编辑 2 E; Q# h+ t2 K0 n& o4 J, f
8 A2 z4 Y4 J6 P% GCurrent Protocols in Stem Cell Biology
7 w$ L/ }8 C5 ?- Z% `Last Update: July 07, 20091 ]) w' _! z6 }" C( S& F: a/ u5 G
Print ISSN: 1941-7322( I. j0 _$ R, [2 K$ l) J$ M
Online ISSN: 1938-8969
! P" p: z% H5 b& f/ x5 {$ c, b% tTable of Contents# f W+ e* ^5 P$ e0 N1 R
Foreword
: A6 Y( @; l4 U9 ]# FPreface
: b. j7 m8 n# t* ~ Chapter 1 Embryonic and Extraembryonic Stem Cells
8 v p; j4 E: P3 p6 O3 n Section A Isolation of Embryonic Stem Cells " \( y9 N. ^3 j- ]: D4 I" \' X
Isolation of Embryonic Stem Cells 4 p; O# i: g ~, l( z) G
UNIT 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells
0 [: V: l; w; x' j3 C! L0 YUNIT 1A.2 Derivation of hESC from Intact Blastocysts
5 k: O% L5 Z8 F( N" t9 dUNIT 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells
- a( X0 g; k$ L( A0 D2 c5 DUNIT 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment & i, S6 w) H+ q# d* _6 }1 Z2 F
Section B Characterization of Embryonic Stem Cells # b, \& K* x2 d1 G" m- ]6 s- ~3 G
UNIT 1B.1 Proteomic Analysis of Pluripotent Stem Cells
" C$ r9 t. v! N) H" e& rUNIT 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and Embryoid Body–Derived Cells Using a High‐Throughput Microarray Platform
: M) ?4 t9 F# [1 T& iUNIT 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells , W |$ V4 L0 S# T+ o; T
UNIT 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency $ D9 h; \. e ?
UNIT 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation ! b. B) r+ u) P o8 g( q/ D
Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells ( N. `. k4 o$ O* R7 V% {. f1 B
Culture and Maintenance of Undifferentiated Embryonic Stem Cells 3 r; m( d7 z& B- w7 ^) ^' D
UNIT 1C.1 Expansion of Human Embryonic Stem Cells In Vitro
8 N. [1 @4 |* J3 |1 X, ^9 @UNIT 1C.2 Defined, Feeder‐Independent Medium for Human Embryonic Stem Cell Culture
" Z2 v. u4 {$ w: m' sUNIT 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells & Q$ s! ~ C5 \0 F
UNIT 1C.4 Culture of Mouse Embryonic Stem Cells
! n7 Z$ n; S# J Y1 }UNIT 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells - a0 F/ J1 o5 J! m) W' V
UNIT 1C.6 Isolation of Human Placental Fibroblasts
5 c9 r; g4 c; [UNIT 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells 3 P7 K' H" \3 N4 ~! f: J4 [
UNIT 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of ROCK Inhibitor " l) c) B: O+ H% u2 l, U) g
Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells 8 [% x, |& l, R- [5 v. m1 j
UNIT 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap & o6 f; B e# D4 z3 T5 z
UNIT 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures
. C1 @& u3 R& `- ? O1 cUNIT 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay % n. g( C: M7 x' r3 A
UNIT 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically Defined Culture Conditions / `) _. f8 Q% `; Q7 ^
UNIT 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells " I; T2 Q+ Z7 {) U3 t- T
Section E Extraembryonic Lineages 7 H8 y3 W. [6 N' a
Isolation of Stem Cells from Extraembryonic Tissues 5 ^' L G. h& q* U! C" C
UNIT 1E.1 Isolation of Human Placenta‐Derived Multipotent Cells and In Vitro Differentiation into Hepatocyte‐Like Cells . c, X6 C7 G' J0 S* B9 e9 \% l
UNIT 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta
1 g$ R! D' I2 P; }9 {( H4 i& FUNIT 1E.3 Isolation of Amniotic Epithelial Stem Cells * Y5 g1 r0 Q M0 m- H
UNIT 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells 0 G, P X; e4 ]2 j
Section F Mesodermal Lineages
* c# q' p" _( PUNIT 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells
4 X. F$ F* g1 ~+ UUNIT 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture with Endoderm in Serum‐Free Medium : \8 R6 ~& G' k4 r
UNIT 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells
) h6 B9 o8 Z; _0 WUNIT 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood , j& p0 }+ t) W; c) w
UNIT 1F.5 Endothelial Differentiation of Embryonic Stem Cells 6 S) @! ]5 C- _0 ^' m* m7 L$ ]
UNIT 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation with Stromal Layers
- g; a. Z6 m# w! @3 j6 _' TUNIT 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors
* w7 B% [% P) o. V G& ^7 BUNIT 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from Human Embryonic Stem Cells ! F# ^/ J% `, F2 [% J" \' ^/ S$ p
Section G Endodermal Lineages
$ n0 A: ~) U+ VUNIT 1G.1 The Differentiation of Distal Lung Epithelium from Embryonic Stem Cells
( |2 z# X1 u+ ^/ p2 zUNIT 1G.2 Pancreas Differentiation of Mouse ES Cells 9 i! \6 Z/ o& ?0 B/ w' I2 q3 q
UNIT 1G.3 Differentiation of Embryonic Stem Cells into Anterior Definitive Endoderm 1 |- P1 p4 q: ^; c4 z
Section H Ectodermal Lineages - B! A) O* E; E& O* m( t
UNIT 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons
% r( ~& C# T4 Q; M Chapter 2 Somatic Stem Cells
, @- l0 ], D6 ]* ^9 x% c Section A Hematopoietic Stem Cells
0 ^. B8 g) c. I. g2 }Hematopoietic Stem Cells
; }1 [ i! J: o& F+ e8 N9 LUNIT 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll‐Paque Density Gradient
* H! a; \; B9 ZUNIT 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood 1 r; N6 z- o: ?/ E' v/ b
UNIT 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood " j; X Z* J# p: @* K j
UNIT 2A.4 Isolation and Assessment of Long‐Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow . {$ f7 K: y2 D/ F* M
UNIT 2A.5 Analysis of the Hematopoietic Stem Cell Niche
; G6 n6 m. w5 i i7 [( qUNIT 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine Embryonic Tissues
" m5 e1 V) [/ G2 sUNIT 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells
) l- Q6 U: P. tUNIT 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells
: s+ @8 g2 }' S' S- ~1 V Section B Non‐Hematopoietic Bone Marrow‐Derived Stem Cells ( U: i7 X6 B* i6 Q2 E
UNIT 2B.1 Isolation and Characterization of Mesoangioblasts from Mouse, Dog, and Human Tissues
( ~/ I: P- M, Z b3 G* j) gUNIT 2B.2 Purification and Culture of Human Blood Vessel–Associated Progenitor Cells
' \ x0 ~- G. IUNIT 2B.3 Isolation, Culture, and Differentiation Potential of Mouse Marrow Stromal Cells ) X1 S4 f( ^0 l+ j
Section C Cardiovascular Stem Cells : T& d. l2 K* v; B" h9 v* @
UNIT 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood
( P1 k0 `% @9 G- o" a) }UNIT 2C.2 Derivation of Epicardium‐Derived Progenitor Cells (EPDCs) from Adult Epicardium [! B3 z! w% ^" P
Section D Neural Stem Cells
3 M _; A: B6 HUNIT 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains % B1 f( L& B( [* R
UNIT 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells 1 H1 g0 c `* K" {8 V2 O. t
UNIT 2D.3 Long‐Term Multilayer Adherent Network (MAN) Expansion, Maintenance, and Characterization, Chemical and Genetic Manipulation, and Transplantation of Human Fetal Forebrain Neural Stem Cells 3 }) O- z! ?% k+ ~9 l( M- o) C
Chapter 3 Cancer Stem Cells ' Z* ^5 P' m3 O( E2 y
UNIT 3.1 Colon Cancer Stem Cells 7 G: b+ d5 S9 z& E1 e
UNIT 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model
6 u4 O/ D }4 y: r" r3 G4 \- X Chapter 4 Manipulation of Potency
2 I% [1 F* N* q y9 L) \3 U& v G Section A iPS Cells : t8 N) o# j4 g2 b& ?$ k( P8 c
UNIT 4A.1 Human iPS Cell Derivation/Reprogramming
) C9 ]3 N! }3 {* R! W" L/ g mUNIT 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells
- s" r# k5 O8 W& G) B% F; y Section B Nuclear Transfer
% l- M5 |5 g" A# \UNIT 4B.1 Heterokaryon‐Based Reprogramming for Pluripotency & Q- _$ s+ F$ l7 |' Q& H
Chapter 5 Genetic Manipulation of Stem Cells 4 C, X3 e3 P3 Y `( \4 J1 Y
Section A Lineage Tracers in Stem Cells
& R" {2 I4 Y7 hUNIT 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma
8 g4 ~$ [5 G4 r7 f( ?UNIT 5A.2 Functional Analysis of Adult Stem Cells Using Cre‐Mediated Lineage Tracing
2 [7 t; z6 t9 m Appendix 1 Useful Information # N9 g4 ^+ y6 a& L# {3 {
APPENDIX 1A Guidelines for the Conduct of Human Embryonic Stem Cell Research
9 I" {) }6 _- Y, I hAPPENDIX 1B ISSCR Guidelines for the Clinical Translation of Stem Cells / P0 P+ L: }1 F5 ?8 _
Appendix 2 Laboratory Equipment # i& f; F2 _$ L
APPENDIX 2 Standard Laboratory Equipment
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