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本帖最后由 细胞海洋 于 2009-8-13 12:01 编辑
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Current Protocols in Stem Cell Biology
4 k. |" L0 A) aLast Update: July 07, 2009/ J5 v# x3 {9 B
Print ISSN: 1941-7322
+ X8 [6 j3 A8 n2 h6 wOnline ISSN: 1938-8969
' S' o) A6 E b* x" b9 n5 cTable of Contents
$ W/ W) Q1 D* w# OForeword
( g! h5 ?) C- \/ g# c% A0 h# FPreface
! D7 g7 u- m* K [1 i" {' I Chapter 1 Embryonic and Extraembryonic Stem Cells
3 n) u- {- U# \& j* ~ Section A Isolation of Embryonic Stem Cells 2 X3 ]4 \( _0 G
Isolation of Embryonic Stem Cells
1 V' v/ p3 J( d; }UNIT 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells
% y. p) M. }, N$ IUNIT 1A.2 Derivation of hESC from Intact Blastocysts
9 ]) a( P& y2 S WUNIT 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells 1 f# M, Q3 i8 U7 q
UNIT 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment 6 U- I( A- S; _2 `1 @
Section B Characterization of Embryonic Stem Cells & _9 u: e+ u# T2 s7 y. ^ d
UNIT 1B.1 Proteomic Analysis of Pluripotent Stem Cells # o. u9 s- h: C
UNIT 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and Embryoid Body–Derived Cells Using a High‐Throughput Microarray Platform . G% R0 t, | a! U
UNIT 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells 0 Q) P) d# e( t6 f
UNIT 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency ) p& Q d) Y$ w1 ^8 k: q
UNIT 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation
& a* g6 ?" d' T Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells # j; l( V9 e, y$ H& b8 A o) G
Culture and Maintenance of Undifferentiated Embryonic Stem Cells
# b% Y0 i5 v% N. w3 ~UNIT 1C.1 Expansion of Human Embryonic Stem Cells In Vitro - R& f: |7 ]( Q) N$ P
UNIT 1C.2 Defined, Feeder‐Independent Medium for Human Embryonic Stem Cell Culture
3 _0 u0 g$ r' P3 F% SUNIT 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells , G3 v+ Z+ W7 _. P
UNIT 1C.4 Culture of Mouse Embryonic Stem Cells ; S; w2 `0 y6 `* V/ _ p7 }
UNIT 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells " g& }2 z2 b! z- @4 A& }( }
UNIT 1C.6 Isolation of Human Placental Fibroblasts
! v& c% C! P# `9 X# d* R7 wUNIT 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells
# ^, t. q" F) A( R# sUNIT 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of ROCK Inhibitor
g7 A! x: G3 C3 P5 }. _( K. b/ d: x1 ] Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells
/ @2 [1 Q; \* r6 _/ B8 |: @7 j( x2 O8 w# `UNIT 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap ( t; }6 T Y+ A' A3 W( |' K% ^
UNIT 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures
2 v. m* @1 g- w9 X* Z6 NUNIT 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay
, k% y" C' C, l' q t( IUNIT 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically Defined Culture Conditions 7 t; b* u7 D p+ Z# Q) p9 }
UNIT 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells 3 T5 C) \+ B5 z3 {9 O3 d" h9 D
Section E Extraembryonic Lineages
" O& x& ?5 W0 D5 F5 e) oIsolation of Stem Cells from Extraembryonic Tissues
0 o% E5 G$ ^9 D7 UUNIT 1E.1 Isolation of Human Placenta‐Derived Multipotent Cells and In Vitro Differentiation into Hepatocyte‐Like Cells
' M8 b) z8 }/ |, C5 F' u- U; QUNIT 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta / c$ y n0 a( r# R
UNIT 1E.3 Isolation of Amniotic Epithelial Stem Cells , S/ R4 M/ H" d6 A8 B5 g+ d! K
UNIT 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells
6 _1 V" k2 D r& J6 K8 t; P Section F Mesodermal Lineages
/ P8 v( F& M* X3 ]- ?1 _UNIT 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells 4 H5 |9 R& W; X0 J0 f- g; y! {. G
UNIT 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture with Endoderm in Serum‐Free Medium . ?- D: h2 i$ ~5 F$ c! p& i, V
UNIT 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells
6 ^) b. P i7 s3 }: [. }, W' hUNIT 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood - n- |1 P3 w6 ^7 w& t, |+ X3 T
UNIT 1F.5 Endothelial Differentiation of Embryonic Stem Cells
) L5 l. R" ?# V9 p# {# g( ?UNIT 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation with Stromal Layers 9 m+ J2 X G/ q7 W, F h
UNIT 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors
4 C0 `# O1 E) g2 z: XUNIT 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from Human Embryonic Stem Cells , y+ R3 ^1 K( }- }4 X/ M. t
Section G Endodermal Lineages
, `( z; l! |" [* `; g& |UNIT 1G.1 The Differentiation of Distal Lung Epithelium from Embryonic Stem Cells 0 G8 I+ r$ f8 v0 u! j' E, N2 H2 S
UNIT 1G.2 Pancreas Differentiation of Mouse ES Cells
2 M! B9 Z8 q1 P" f, {1 `UNIT 1G.3 Differentiation of Embryonic Stem Cells into Anterior Definitive Endoderm 0 ?/ E$ B9 R4 E! L# \ s
Section H Ectodermal Lineages
4 H. z1 p3 U7 m) G, S5 KUNIT 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons
" d' R4 f$ V A; O$ d: c5 W6 a3 c Chapter 2 Somatic Stem Cells ' D# S O% Z; R3 e- |4 |
Section A Hematopoietic Stem Cells 7 H' h! h- F7 S8 s. L1 {
Hematopoietic Stem Cells
0 ^ o u1 x2 }6 X% [' K, r1 z* lUNIT 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll‐Paque Density Gradient * y; W- P+ [: s, f
UNIT 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood ! M- F. ]6 ~" s/ K
UNIT 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood
, M" D, e# e& |8 ]* Z8 qUNIT 2A.4 Isolation and Assessment of Long‐Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow " S# b" M% i9 ?- }' H, }9 Q
UNIT 2A.5 Analysis of the Hematopoietic Stem Cell Niche ' }$ U# [/ K- Z! f8 Q0 s. }9 E
UNIT 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine Embryonic Tissues
) N5 Q/ D, k T! K7 i9 N! |$ \( F+ TUNIT 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells % e) F: H% l8 _1 l' m1 f8 i
UNIT 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells 4 [/ |8 T% z) D+ f; r! ^9 |- M
Section B Non‐Hematopoietic Bone Marrow‐Derived Stem Cells : X8 f% }: D5 b7 [ p% m7 }+ r
UNIT 2B.1 Isolation and Characterization of Mesoangioblasts from Mouse, Dog, and Human Tissues ' _1 a6 A7 d1 m
UNIT 2B.2 Purification and Culture of Human Blood Vessel–Associated Progenitor Cells % ?7 A* g6 p2 X' ~
UNIT 2B.3 Isolation, Culture, and Differentiation Potential of Mouse Marrow Stromal Cells
5 t9 Q' e v w4 U9 R) s Section C Cardiovascular Stem Cells
0 Y! l/ _& N7 z* UUNIT 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood
5 @/ {; v4 E* O4 @. X9 PUNIT 2C.2 Derivation of Epicardium‐Derived Progenitor Cells (EPDCs) from Adult Epicardium & p+ ~4 S g P7 |& |1 @- k
Section D Neural Stem Cells : i% E8 b7 X" E! c
UNIT 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains 7 T1 `, p n8 A5 V
UNIT 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells
" [; o: e& v" b B1 L2 f8 iUNIT 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 7 O" I% k d# i+ ?
Chapter 3 Cancer Stem Cells
/ X. a0 ~) I$ j' q% F% h' }( v) z5 sUNIT 3.1 Colon Cancer Stem Cells
8 x5 L5 U- O3 V( r5 c% OUNIT 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model 5 Q* \* f2 F# B% h5 z- ^, B( E3 p
Chapter 4 Manipulation of Potency 2 h: v, }1 @: y6 R
Section A iPS Cells
' m5 V+ t- w# Z% j: a3 {UNIT 4A.1 Human iPS Cell Derivation/Reprogramming & n! A; T3 P" y3 v& B/ d
UNIT 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells
7 N- z- U, p# n- i6 i Q6 K9 Q Section B Nuclear Transfer * G i, \, @ F# F% z x( ]
UNIT 4B.1 Heterokaryon‐Based Reprogramming for Pluripotency
/ y/ C: X5 l# A& O; y8 m, E1 K4 o Chapter 5 Genetic Manipulation of Stem Cells
. s( q4 W+ W/ N Section A Lineage Tracers in Stem Cells
+ W7 o& x7 Q3 U+ T% qUNIT 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma ( M0 w4 O! ^( D* I0 h; q4 V/ J
UNIT 5A.2 Functional Analysis of Adult Stem Cells Using Cre‐Mediated Lineage Tracing . [2 v" \) o3 f `
Appendix 1 Useful Information % e0 `1 \9 n+ G; ~
APPENDIX 1A Guidelines for the Conduct of Human Embryonic Stem Cell Research
( R; W* [7 ^, A# @; A& q5 ]7 eAPPENDIX 1B ISSCR Guidelines for the Clinical Translation of Stem Cells 0 u F* ]6 |* R5 @6 {8 x5 }$ S1 Z
Appendix 2 Laboratory Equipment : r3 U3 O: a5 C+ O4 q; c4 X" u$ g
APPENDIX 2 Standard Laboratory Equipment v. i$ Z2 `0 _+ x) T3 K
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