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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
3 x- A- z* ]) m: b' X1 M3 ~9 g7 \; Z( \Last Update: July 07, 2009
5 P) |$ k: R# Z9 L9 a& u dPrint ISSN: 1941-7322; ?, y7 v7 R+ u4 i! u9 N' h, e/ O
Online ISSN: 1938-89696 h/ \. c) u( q
Table of Contents% o" k! A- T% L4 [9 J% U# e. z
Foreword 7 Z+ |! z, q1 r7 L8 q- m! ~
Preface G I# R/ R G( I
Chapter 1 Embryonic and Extraembryonic Stem Cells
, @6 R! b6 i D8 d4 S9 j Section A Isolation of Embryonic Stem Cells
" y4 @0 \5 n& e8 C( O+ IIsolation of Embryonic Stem Cells
) @+ j8 n0 [$ x# ? D NUNIT 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells
2 u& j7 L7 m1 ?6 Q& b- cUNIT 1A.2 Derivation of hESC from Intact Blastocysts
3 [& R9 g8 G+ I2 s! x1 u4 b2 tUNIT 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells : l+ v' D* x5 S+ h- f5 q8 Y
UNIT 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment
% _; d" V2 R0 S# ^ Section B Characterization of Embryonic Stem Cells " S2 A( c+ P) o, l! W
UNIT 1B.1 Proteomic Analysis of Pluripotent Stem Cells
7 b i6 ~% s; D, k! `% DUNIT 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and Embryoid Body–Derived Cells Using a High‐Throughput Microarray Platform
8 D o3 W* ~$ K' n% r& E* FUNIT 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells
7 S; S" W( u8 G- L5 oUNIT 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency
- \4 J9 o$ k! C+ c# M6 SUNIT 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation , T5 S& \, R4 M" D8 u; H8 b
Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells
! V R R9 N w# s. ~Culture and Maintenance of Undifferentiated Embryonic Stem Cells
' ?! K* {, m/ P* r/ s. j7 f. IUNIT 1C.1 Expansion of Human Embryonic Stem Cells In Vitro ; c9 F) r; I+ |6 m! U K
UNIT 1C.2 Defined, Feeder‐Independent Medium for Human Embryonic Stem Cell Culture
" m- q6 v/ \6 x' {5 n3 NUNIT 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells ! u* s/ I1 N9 j# W3 T K
UNIT 1C.4 Culture of Mouse Embryonic Stem Cells ) W" t; m; h, h0 z# _
UNIT 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells
: b) e$ N) Z+ y3 Q. fUNIT 1C.6 Isolation of Human Placental Fibroblasts
2 c8 t6 C6 T' b9 e; WUNIT 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells
: t& g X0 H6 s6 f- RUNIT 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of ROCK Inhibitor
% }( Y3 ^* R i8 ^; a8 B! K3 z Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells & s' x G/ k- a. F
UNIT 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap
" O# U/ ~6 B6 \& U& s9 tUNIT 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures
5 n2 e. k7 a0 C$ W; VUNIT 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay
% ~0 c$ `6 m1 _% Q) \UNIT 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically Defined Culture Conditions $ v4 `, F* Y/ e6 _' g0 N P
UNIT 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells ) r. B0 t, _$ n. g/ @2 z+ s; _$ o8 `
Section E Extraembryonic Lineages ! p8 U3 {& R7 }5 a! w' x5 C
Isolation of Stem Cells from Extraembryonic Tissues & A. I! N% ^+ C6 A
UNIT 1E.1 Isolation of Human Placenta‐Derived Multipotent Cells and In Vitro Differentiation into Hepatocyte‐Like Cells 3 G6 H; r. K( o0 L
UNIT 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta
( m# D) e5 f+ }- V; @UNIT 1E.3 Isolation of Amniotic Epithelial Stem Cells , R* w7 l! g6 x: _0 p. V8 v& @
UNIT 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells f: a) l" B& O$ D6 t8 I5 }
Section F Mesodermal Lineages
5 z! p( R) t5 C. W) R% m# U8 W; hUNIT 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells
9 Q# \1 }6 g% G: I P9 i/ AUNIT 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture with Endoderm in Serum‐Free Medium
0 [, B! `! {# |8 S# I& fUNIT 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells 8 b- ~$ Q$ A: H( m3 V
UNIT 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood
5 O: q/ Z& }/ `- z6 GUNIT 1F.5 Endothelial Differentiation of Embryonic Stem Cells % z. }" T" V4 p* \
UNIT 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation with Stromal Layers
% S# G* ^+ h: o/ i7 ?UNIT 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors 4 ~$ w. e- w3 I# V3 A
UNIT 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from Human Embryonic Stem Cells ' K: R6 D1 N) z: z$ ^) N
Section G Endodermal Lineages
, w0 n6 N) O$ ~) mUNIT 1G.1 The Differentiation of Distal Lung Epithelium from Embryonic Stem Cells
+ f" ~! x5 A2 t$ ?" Z# ~4 sUNIT 1G.2 Pancreas Differentiation of Mouse ES Cells 6 O8 I( F7 o6 @4 M) X2 c/ B$ h- G
UNIT 1G.3 Differentiation of Embryonic Stem Cells into Anterior Definitive Endoderm
' e& ]% j) h% k( [5 M' @ Section H Ectodermal Lineages : u. c0 Q! h n3 a4 G0 r9 I7 @6 m
UNIT 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons & ]( _4 y, A5 R) F, `9 q8 ]$ z# E
Chapter 2 Somatic Stem Cells
9 e' m! _( q% Q1 h0 U Section A Hematopoietic Stem Cells $ |9 j& H( \% [
Hematopoietic Stem Cells 5 Y, P1 L' O6 m, _8 n
UNIT 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll‐Paque Density Gradient 1 W, ^- r( W U% ]
UNIT 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood
: c" y+ `: \0 ]$ ?0 |UNIT 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood 3 n* C' o" i1 u: U
UNIT 2A.4 Isolation and Assessment of Long‐Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow 8 x3 U9 ~' J" f9 L
UNIT 2A.5 Analysis of the Hematopoietic Stem Cell Niche & C+ c0 O: M& O1 O
UNIT 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine Embryonic Tissues 8 I( G2 M, J4 [
UNIT 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells
& }& w( T0 Z- n! W6 UUNIT 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells
/ n; Y/ C; h i$ a3 v Section B Non‐Hematopoietic Bone Marrow‐Derived Stem Cells - ]: B2 \; P. r9 B; V8 R8 X) |& ~
UNIT 2B.1 Isolation and Characterization of Mesoangioblasts from Mouse, Dog, and Human Tissues 8 v( j& s4 n/ D1 k! L+ }
UNIT 2B.2 Purification and Culture of Human Blood Vessel–Associated Progenitor Cells
) l, b' y& q, Z3 ]9 s: wUNIT 2B.3 Isolation, Culture, and Differentiation Potential of Mouse Marrow Stromal Cells ' a. P! a$ |$ ]
Section C Cardiovascular Stem Cells
0 O }4 D6 c A& G$ f" g UUNIT 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood
" Q7 [! E) R1 d8 O' G3 EUNIT 2C.2 Derivation of Epicardium‐Derived Progenitor Cells (EPDCs) from Adult Epicardium . N; t4 d+ r, A4 f$ a; m
Section D Neural Stem Cells
, W, q8 X& P8 C$ {UNIT 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains
# o' v7 g! ~" H3 J% `: FUNIT 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells 5 b6 Y0 b1 {9 `8 m
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 ! D+ K! ]5 G1 f Q
Chapter 3 Cancer Stem Cells 9 n: `5 c! p3 L
UNIT 3.1 Colon Cancer Stem Cells
6 s( L/ \2 g$ C3 l' A" k3 CUNIT 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model ! Z! B; G2 G5 ~) \* _. c- F
Chapter 4 Manipulation of Potency
9 G7 W: J% z" T5 C Section A iPS Cells ! \5 V+ B# t: m* k4 X
UNIT 4A.1 Human iPS Cell Derivation/Reprogramming $ _5 [7 Z0 I) a
UNIT 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells 8 h3 q6 @3 \9 a* u0 |+ @0 P
Section B Nuclear Transfer ( a1 j0 J4 E8 t: x. \% H1 |7 j6 R
UNIT 4B.1 Heterokaryon‐Based Reprogramming for Pluripotency
* I- ]$ [% O& H% [9 f; e6 O% w Chapter 5 Genetic Manipulation of Stem Cells
2 ]( E) f1 Y$ V9 G; Y0 J Section A Lineage Tracers in Stem Cells ( s- A2 S4 w* g$ Z7 P& }& j9 @' \
UNIT 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma
( R3 _. K* u8 p' h) U7 Z. AUNIT 5A.2 Functional Analysis of Adult Stem Cells Using Cre‐Mediated Lineage Tracing $ m2 H7 ?) i, M5 P7 O* V6 d/ {1 p
Appendix 1 Useful Information * m) [' [. g# W- J" Y- X
APPENDIX 1A Guidelines for the Conduct of Human Embryonic Stem Cell Research % L# ]- l5 T+ Q7 m$ ^% q$ A
APPENDIX 1B ISSCR Guidelines for the Clinical Translation of Stem Cells
, o2 u* `4 K5 @" ?# D0 J- Q Appendix 2 Laboratory Equipment
1 v. a* U; |8 G/ u! M- o2 ^. XAPPENDIX 2 Standard Laboratory Equipment - _) x0 |- J& z4 S# l6 U. i
先谢谢啦! |
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