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本帖最后由 细胞海洋 于 2009-8-13 12:01 编辑 % t/ t9 ? m, V" }
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Current Protocols in Stem Cell Biology
' g7 p4 C: ?, a" Q* M7 TLast Update: July 07, 2009( t' V; g+ h! ~. ^$ T
Print ISSN: 1941-7322
; }; o! m/ e& P8 |- dOnline ISSN: 1938-8969
$ p# `( H; F; f# \0 ^Table of Contents. L0 j7 u5 _# Y$ Y
Foreword
; u+ ?$ L: y: K3 xPreface 6 C6 t3 C6 p- J, S) g3 |
Chapter 1 Embryonic and Extraembryonic Stem Cells w7 m* C$ a+ @$ E& N1 \
Section A Isolation of Embryonic Stem Cells
5 t0 }* z" ]. ~* N. S6 lIsolation of Embryonic Stem Cells / }$ F% G8 b( `1 a+ R/ v
UNIT 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells
$ o( @) e# ]5 P5 \" I$ ]% f- \# HUNIT 1A.2 Derivation of hESC from Intact Blastocysts ( J& u% W5 E: H5 P( S) k: Q
UNIT 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells ! M, I- _* ~: v% R
UNIT 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment N& [# Y- M( W0 A( ]
Section B Characterization of Embryonic Stem Cells
9 W* d+ R% W$ J4 {- ~ D4 P+ PUNIT 1B.1 Proteomic Analysis of Pluripotent Stem Cells
4 b$ _2 b- l5 x0 L4 QUNIT 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and Embryoid Body–Derived Cells Using a High‐Throughput Microarray Platform
) ? v- C% Q/ C5 `9 {UNIT 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells * u2 ]) u& R. q b0 K C
UNIT 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency ( L- N3 y" P/ A# _" C! f- f
UNIT 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation
8 p% Y, ?! ^- O, H# _/ S U' C) j Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells
7 s S% ^/ ]* Z8 G& vCulture and Maintenance of Undifferentiated Embryonic Stem Cells & K5 f& m1 y) m( {: ~. x( t: c9 J
UNIT 1C.1 Expansion of Human Embryonic Stem Cells In Vitro
+ B c3 A1 a- _3 RUNIT 1C.2 Defined, Feeder‐Independent Medium for Human Embryonic Stem Cell Culture
$ s1 a6 c4 O$ JUNIT 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells $ S9 M o- F# l
UNIT 1C.4 Culture of Mouse Embryonic Stem Cells
@: d9 e& p3 c6 Q1 E$ G o1 o. D6 q4 QUNIT 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells 3 c% g% }8 L7 S1 J% Q! V
UNIT 1C.6 Isolation of Human Placental Fibroblasts
1 j# o. d& e3 n8 N4 TUNIT 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells
% W/ g, ?3 J/ Y7 L: K/ u( P2 {% }; I$ ~& [UNIT 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of ROCK Inhibitor , a) B/ ^9 \, C3 O9 }/ K5 w! G1 S- e
Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells
# [+ l" |! x/ H7 c2 Q& e# ZUNIT 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap 0 |- b$ N8 l% Y$ E* o
UNIT 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures
" O* O1 K' o/ n/ t# E! @UNIT 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay
0 ?: n) h, j3 F+ aUNIT 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically Defined Culture Conditions . L, X! D/ ?. T
UNIT 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells - S/ I) f7 Z) c
Section E Extraembryonic Lineages
7 W9 y ^' v% }" r3 Q; G% g [Isolation of Stem Cells from Extraembryonic Tissues * n) q6 i1 y) B1 Y9 c
UNIT 1E.1 Isolation of Human Placenta‐Derived Multipotent Cells and In Vitro Differentiation into Hepatocyte‐Like Cells ! M$ @. s2 L9 d8 O# M/ d
UNIT 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta
" G, _2 f. R- WUNIT 1E.3 Isolation of Amniotic Epithelial Stem Cells
& ?. E& d' x: w& h: z& sUNIT 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells
# {. h) ?& z# h L5 f' }0 L Section F Mesodermal Lineages M* D0 d7 z# x; K& D
UNIT 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells
5 H; J* X- l9 o& |0 ?UNIT 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture with Endoderm in Serum‐Free Medium " j/ j7 F; ?# o& n1 [( F9 K9 y
UNIT 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells
+ k6 N6 _& h% J5 C3 w& Q VUNIT 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood
2 m8 l0 g, O: l" v) J+ S9 E- tUNIT 1F.5 Endothelial Differentiation of Embryonic Stem Cells ) q# e% h) |$ {! b
UNIT 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation with Stromal Layers " }- B# M, f9 P
UNIT 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors 0 s, l1 p, m$ G3 z) l
UNIT 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from Human Embryonic Stem Cells
Z$ i5 M* H5 f. d# r Section G Endodermal Lineages
( D$ z# t( A: o3 |* KUNIT 1G.1 The Differentiation of Distal Lung Epithelium from Embryonic Stem Cells
1 y+ T0 X4 t: lUNIT 1G.2 Pancreas Differentiation of Mouse ES Cells
' l7 `, V. I2 t, v7 }1 }8 L8 ?UNIT 1G.3 Differentiation of Embryonic Stem Cells into Anterior Definitive Endoderm
6 N: o* c/ W. | Section H Ectodermal Lineages 5 H9 I) t. ~) V- u+ n% S0 ]- \
UNIT 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons
8 n0 A6 A7 L S* Z Chapter 2 Somatic Stem Cells
0 }) |9 c, ~. ] Section A Hematopoietic Stem Cells
+ \; O: s- V1 IHematopoietic Stem Cells 2 ~/ R2 b3 i$ k7 y( f% U6 g2 q
UNIT 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll‐Paque Density Gradient
" _0 W& E- u! OUNIT 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood ) E7 L9 C6 \4 z. X
UNIT 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood . J8 Z( t+ F1 _2 }' `8 Q5 ]# W3 a
UNIT 2A.4 Isolation and Assessment of Long‐Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow 9 M+ Q" r" e# h3 l* J. K Z! ]
UNIT 2A.5 Analysis of the Hematopoietic Stem Cell Niche
. G4 F5 H" {( PUNIT 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine Embryonic Tissues
3 ?/ N/ F- d* k8 k4 G% vUNIT 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells
$ E, c7 C( Z3 F- b2 j- t( M- PUNIT 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells 4 D. ?8 F. `) a" ?7 d) L
Section B Non‐Hematopoietic Bone Marrow‐Derived Stem Cells
7 [5 O% d& j, K6 i3 @. Y. v! i! uUNIT 2B.1 Isolation and Characterization of Mesoangioblasts from Mouse, Dog, and Human Tissues
% A6 P! \4 {/ a+ Q4 q. d5 VUNIT 2B.2 Purification and Culture of Human Blood Vessel–Associated Progenitor Cells , U z& x2 _ D
UNIT 2B.3 Isolation, Culture, and Differentiation Potential of Mouse Marrow Stromal Cells
# @% ^' Y/ ]9 p8 D- L Section C Cardiovascular Stem Cells " J+ N6 P+ V" i! l* g
UNIT 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood
* L- I u5 ]& x( bUNIT 2C.2 Derivation of Epicardium‐Derived Progenitor Cells (EPDCs) from Adult Epicardium
- l# g; j% V, w Section D Neural Stem Cells % a, \" k9 y4 _8 }8 f
UNIT 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains
& j% Q) n4 r# vUNIT 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells
1 o" O) [! G+ LUNIT 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 m4 N2 P7 ?. r1 k) A Chapter 3 Cancer Stem Cells
; H. N2 }3 c% S5 W' o7 uUNIT 3.1 Colon Cancer Stem Cells ! P8 U: q- C7 ~) l. p; }+ ^6 n
UNIT 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model
6 p* ~; q1 \9 ~" ?( e. _3 J Chapter 4 Manipulation of Potency 5 t' O4 o$ Y) I" b9 I' E
Section A iPS Cells + k- [6 q$ u, N% p5 S+ v5 C
UNIT 4A.1 Human iPS Cell Derivation/Reprogramming
& z* S9 n: }: j/ @UNIT 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells ! v9 o4 s; K, }6 Y8 X/ m
Section B Nuclear Transfer
/ b. [/ Z- D8 y1 s% g$ Z$ R: tUNIT 4B.1 Heterokaryon‐Based Reprogramming for Pluripotency K- g) D. ~% e; J8 Y( _0 B
Chapter 5 Genetic Manipulation of Stem Cells
`* \4 ^; R7 _! j Section A Lineage Tracers in Stem Cells 7 f: Z- x* _* x6 T7 {
UNIT 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma 4 V# U) h& C8 H! P% l
UNIT 5A.2 Functional Analysis of Adult Stem Cells Using Cre‐Mediated Lineage Tracing 4 F* q# K# s# }
Appendix 1 Useful Information
& J) O( P5 L) k; e% Q: M; j" g( E$ YAPPENDIX 1A Guidelines for the Conduct of Human Embryonic Stem Cell Research 2 d2 O) I) U# P9 W6 y/ A: S+ v
APPENDIX 1B ISSCR Guidelines for the Clinical Translation of Stem Cells
2 k9 ~' W2 E- h% f! Z) } Appendix 2 Laboratory Equipment 5 v# R, M& X9 n
APPENDIX 2 Standard Laboratory Equipment
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