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本帖最后由 细胞海洋 于 2009-8-13 12:01 编辑 ; t, H3 R* _( P7 W
$ B+ h; @, [ t7 S5 zCurrent Protocols in Stem Cell Biology+ P7 i- ~9 h! i4 ^- ~3 d
Last Update: July 07, 2009
' l# j& t5 M* U+ ~2 iPrint ISSN: 1941-7322% O+ v& D4 M. k
Online ISSN: 1938-8969
- }7 o8 e5 U% x& F. _( |! j* uTable of Contents6 j* l$ r, ]. u1 k( ~' H
Foreword
3 q) Z8 X5 O$ g4 {% KPreface
3 f3 V$ }% d- Q Chapter 1 Embryonic and Extraembryonic Stem Cells
8 ?5 z& d# z5 r- o ~ Section A Isolation of Embryonic Stem Cells
. w+ o6 E6 t0 AIsolation of Embryonic Stem Cells
6 H0 Y$ a7 F- c. Y) pUNIT 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells
7 G8 p+ X1 k0 F2 h3 W9 [( v! }UNIT 1A.2 Derivation of hESC from Intact Blastocysts
6 x/ H8 }3 s; A2 l9 R. L( O/ r; RUNIT 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells / E% E i4 [; Y- B/ |
UNIT 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment & r& T3 q9 C9 b2 t4 x+ H' Y
Section B Characterization of Embryonic Stem Cells
# D# c# }2 x }! j1 C3 u/ cUNIT 1B.1 Proteomic Analysis of Pluripotent Stem Cells 6 z) M( E) T3 w; K+ m
UNIT 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and Embryoid Body–Derived Cells Using a High‐Throughput Microarray Platform & f' v# u4 D/ B1 q1 G; L6 u. m$ b
UNIT 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells 3 `/ ?, E4 _) k9 ]& i5 Y
UNIT 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency 7 l* m7 G; S O# }
UNIT 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation
8 `; o5 `# e$ j3 l6 J Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells
& o+ X, `/ v, y( }9 `% |6 R [Culture and Maintenance of Undifferentiated Embryonic Stem Cells 1 h1 \7 R. p: k# e! S
UNIT 1C.1 Expansion of Human Embryonic Stem Cells In Vitro " p! `& g4 @4 r5 B) V
UNIT 1C.2 Defined, Feeder‐Independent Medium for Human Embryonic Stem Cell Culture " v* A( Z; k H1 k4 C) V
UNIT 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells 3 r8 q7 p2 p* U$ {0 g( x3 {- ]% t
UNIT 1C.4 Culture of Mouse Embryonic Stem Cells
- Q% U0 c6 t; o* r8 W8 SUNIT 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells 1 R+ U% ~* q, W2 F2 X7 F
UNIT 1C.6 Isolation of Human Placental Fibroblasts ( g& w8 Y5 P- i
UNIT 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells
2 u7 \6 w' G; X$ S' W# |" i) K/ \! VUNIT 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of ROCK Inhibitor
6 ?" h+ D$ B) O: p- a3 D Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells
% R5 {6 V/ `- _+ T% nUNIT 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap $ N' c3 ]0 J7 r7 {4 S
UNIT 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures ! V b5 J# @8 S; ^- d7 O6 S$ t2 D
UNIT 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay : {& o; I' r, P2 X9 v8 l$ \, w
UNIT 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically Defined Culture Conditions
' v! ^7 S. ^5 r" W s$ I; ]) _3 h( sUNIT 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells - O2 W. o- P: b" ]6 h
Section E Extraembryonic Lineages
% {1 L* s/ U* q3 c, _1 S6 ?2 BIsolation of Stem Cells from Extraembryonic Tissues
4 @# F" a1 O1 P( x5 H1 TUNIT 1E.1 Isolation of Human Placenta‐Derived Multipotent Cells and In Vitro Differentiation into Hepatocyte‐Like Cells
2 o/ H5 d# V7 g! D; i7 H8 ~UNIT 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta ; A J* X9 I. w( o( @1 f
UNIT 1E.3 Isolation of Amniotic Epithelial Stem Cells ; q8 U9 r: ~" L8 z) n+ T3 b( ^! Q
UNIT 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells
) a: H, n$ ^5 T Section F Mesodermal Lineages ' V' B/ R1 z$ x8 E
UNIT 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells # ~' W ]: T# @7 I6 ~7 A7 ~
UNIT 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture with Endoderm in Serum‐Free Medium
+ u' y4 y+ U" g* v3 t( AUNIT 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells
* ^7 }# a+ ~) Y0 E1 JUNIT 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood
* F, R) B7 t; M5 e) EUNIT 1F.5 Endothelial Differentiation of Embryonic Stem Cells
, L3 [5 J) C! \% c- I! U7 ]UNIT 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation with Stromal Layers 8 P! }" Q. a2 |" ?, C
UNIT 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors
7 H+ F i3 ]5 T1 X# b) JUNIT 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from Human Embryonic Stem Cells " g) V' |; Q7 Z& F" o6 H
Section G Endodermal Lineages 2 M3 q; L, g/ b& d3 m( T) I
UNIT 1G.1 The Differentiation of Distal Lung Epithelium from Embryonic Stem Cells % ]% z9 \5 r( @5 G7 W4 A% O
UNIT 1G.2 Pancreas Differentiation of Mouse ES Cells : M) Q; t1 ~6 ^+ g- t( M1 y
UNIT 1G.3 Differentiation of Embryonic Stem Cells into Anterior Definitive Endoderm
2 P, U6 I6 ?7 G4 f! [. |! z$ Y Section H Ectodermal Lineages
: j* b1 |6 F( A8 P) G- @% CUNIT 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons
W/ L% T6 ^+ U! ?' L' Y) z' M Chapter 2 Somatic Stem Cells ! g! h7 ~" A' z0 Y7 ~- [6 X: `
Section A Hematopoietic Stem Cells ! q8 a" c: U, |" N4 s/ A; F5 l, D+ |
Hematopoietic Stem Cells
3 C; N J- L7 r3 d8 m3 u' d/ GUNIT 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll‐Paque Density Gradient % Y! A& t3 C9 u, l1 t
UNIT 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood 3 X/ \7 _1 [! p% q9 [5 m) ~
UNIT 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood 9 _! K) n; B9 R! O& G# n7 W! L
UNIT 2A.4 Isolation and Assessment of Long‐Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow 5 a9 G0 P/ B% H; s1 b6 c6 y
UNIT 2A.5 Analysis of the Hematopoietic Stem Cell Niche
% m& o# q) ^/ p; h' x# FUNIT 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine Embryonic Tissues
+ b. N) J# B# w% t* I; WUNIT 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells
8 s" W' d+ m& g3 g, k* U9 `UNIT 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells
% \+ g3 Y+ T' F U, p4 q% R Section B Non‐Hematopoietic Bone Marrow‐Derived Stem Cells
9 |' |1 h. z4 d# A0 a6 |6 oUNIT 2B.1 Isolation and Characterization of Mesoangioblasts from Mouse, Dog, and Human Tissues
$ k$ _1 x6 J. C: o8 C9 F3 m4 Y9 GUNIT 2B.2 Purification and Culture of Human Blood Vessel–Associated Progenitor Cells
8 V0 t) k! X) V5 d# J9 JUNIT 2B.3 Isolation, Culture, and Differentiation Potential of Mouse Marrow Stromal Cells
: A$ [9 ^. L6 G5 m3 S/ Z Section C Cardiovascular Stem Cells 0 n$ H' Y, p+ l9 p9 p( d3 w/ `9 |
UNIT 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood . O3 C9 `5 O6 Q7 c: H5 n
UNIT 2C.2 Derivation of Epicardium‐Derived Progenitor Cells (EPDCs) from Adult Epicardium
9 ]" ], T0 k- s Section D Neural Stem Cells
2 y. Y3 K. Y# G8 T: }0 _UNIT 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains
$ j3 e3 A$ W7 M) g3 @UNIT 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells
" Z6 U3 m6 S' F; Y: b& 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 , [& {! Q, }# U6 N
Chapter 3 Cancer Stem Cells
4 {* K4 v+ V, IUNIT 3.1 Colon Cancer Stem Cells
3 J2 k/ m7 }7 h$ K, eUNIT 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model
* y0 _' e6 C3 ^" s/ m6 I' w Chapter 4 Manipulation of Potency ( I3 O1 Y! D. [" P$ N
Section A iPS Cells + P( Y$ b: I# N' b. o$ z
UNIT 4A.1 Human iPS Cell Derivation/Reprogramming
: s5 ~2 k6 G# {5 l1 `UNIT 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells ! n+ q8 A' H8 c1 B' Q! y9 r
Section B Nuclear Transfer 9 Z9 ~/ a& p' d3 _! b0 g& b
UNIT 4B.1 Heterokaryon‐Based Reprogramming for Pluripotency
8 a" I |6 A* `5 Y J Chapter 5 Genetic Manipulation of Stem Cells
: g" E. i9 _* V Section A Lineage Tracers in Stem Cells
7 V. Z, x) ^) L3 F+ {0 UUNIT 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma ! k7 k, F; z3 n+ a8 \; n' o
UNIT 5A.2 Functional Analysis of Adult Stem Cells Using Cre‐Mediated Lineage Tracing - K+ F! {1 u! ~3 D% X" l
Appendix 1 Useful Information 1 R7 C3 N r6 v5 s+ v9 G
APPENDIX 1A Guidelines for the Conduct of Human Embryonic Stem Cell Research + P% u3 a/ U9 u1 M
APPENDIX 1B ISSCR Guidelines for the Clinical Translation of Stem Cells
: p: c8 X0 ?" H+ Q* j( m2 m Appendix 2 Laboratory Equipment : L S ?: L* K& A4 a
APPENDIX 2 Standard Laboratory Equipment
: S) {% I g* }; b先谢谢啦! |
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