求助current protocols in stem cell biology

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Current Protocols in Stem Cell Biology/ P) d7 M0 `( f3 G
Last Update: July 07, 2009
- }- u* Z9 I5 z& B$ HPrint ISSN: 1941-7322
' q- V8 y8 \6 n! k6 cOnline ISSN: 1938-8969
2 \. i- J, E! L% C" d3 n5 o, BTable of Contents
* V# N! v9 }  H5 V2 h% CForeword ! [- S0 A9 r2 S' d4 Z( h% T
Preface 2 n8 V$ ^, b: z% J9 z
Chapter 1 Embryonic and Extraembryonic Stem Cells $ B. F& H: V4 M. c8 l5 m
Section A Isolation of Embryonic Stem Cells 5 ~3 b0 D" l- H* Z- K
Isolation of Embryonic Stem Cells + d0 {" K6 N; l  O
UNIT 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells   Z" B+ ]( l( D0 ~; p1 x* ~% q
UNIT 1A.2 Derivation of hESC from Intact Blastocysts
  i5 v- T2 E: Y' w& s; F- n  sUNIT 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells
' O0 O0 z( ^- ^8 vUNIT 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment
/ c" \  }& w# V8 L, y; z Section B Characterization of Embryonic Stem Cells 5 U( g$ V. r8 H' t
UNIT 1B.1 Proteomic Analysis of Pluripotent Stem Cells : ]+ e% h/ d, V# G6 E) F, m2 a. s; d
UNIT 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and Embryoid Body–Derived Cells Using a High‐Throughput Microarray Platform
$ Q* f, m/ |# k7 K/ d* R( oUNIT 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells
3 Z1 z9 P; w% OUNIT 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency
+ T, Y" O4 y8 s6 L, VUNIT 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation 5 r. \) s( w/ ]0 _0 t0 p
Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells
2 Z" X# y) y7 `. R3 A9 eCulture and Maintenance of Undifferentiated Embryonic Stem Cells ; C. W, S9 g: ?
UNIT 1C.1 Expansion of Human Embryonic Stem Cells In Vitro ) w; ?( u) J& \. w# O& {% y
UNIT 1C.2 Defined, Feeder‐Independent Medium for Human Embryonic Stem Cell Culture
7 l$ N& a- w& r% M  n2 YUNIT 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells
  M, n+ A9 `  @$ r4 B( j5 S, iUNIT 1C.4 Culture of Mouse Embryonic Stem Cells
" Y6 s8 t7 l+ q( X) \2 cUNIT 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells * T6 q/ a- R2 c! Y4 Q1 ]4 |
UNIT 1C.6 Isolation of Human Placental Fibroblasts
* w  h, l- T3 @; I' jUNIT 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells
1 X4 o8 K  u4 X2 T  g* b$ b' N9 {- xUNIT 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of ROCK Inhibitor " O2 e: g' w6 I$ n1 ]+ M
Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells . T: Q0 \  D1 |% X2 \
UNIT 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap 4 B, @- o0 r' n  f2 o2 @! \
UNIT 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures ) E7 C9 L; m  \  G
UNIT 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay
5 r, }1 [2 P' ?" k% {& DUNIT 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically Defined Culture Conditions ( R8 X- C- X3 P" X- Z! U
UNIT 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells : k4 T0 _5 q0 Y6 l1 P$ K
Section E Extraembryonic Lineages
- {) t7 j5 o! L& Q7 lIsolation of Stem Cells from Extraembryonic Tissues - n' J: N- N# O3 K- i6 j! o' P
UNIT 1E.1 Isolation of Human Placenta‐Derived Multipotent Cells and In Vitro Differentiation into Hepatocyte‐Like Cells
  T, D, z. V+ J2 u# t5 wUNIT 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta
( ]  i, S$ N/ S2 \; |+ uUNIT 1E.3 Isolation of Amniotic Epithelial Stem Cells
- q# K6 v6 c$ S6 eUNIT 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells
; ^4 q, d# {8 y0 D2 M Section F Mesodermal Lineages + h2 L5 ]/ o' c7 x# q
UNIT 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells
. w+ t0 M2 e; _7 p( B% \& XUNIT 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture with Endoderm in Serum‐Free Medium
1 h- f" _, |( ^8 T& }$ RUNIT 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells ! o- P" v3 A8 P+ S& r2 ?3 d) c
UNIT 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood 1 o6 |! J; f/ P, `
UNIT 1F.5 Endothelial Differentiation of Embryonic Stem Cells 3 m0 W4 v2 f* r# B% S
UNIT 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation with Stromal Layers
+ Q6 l5 [- O! r6 dUNIT 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors 4 ?5 ?- F# Y, [% x3 o& `
UNIT 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from Human Embryonic Stem Cells
+ e; X' F) f/ R. k( Q Section G Endodermal Lineages " p# Q6 H) p$ p$ a
UNIT 1G.1 The Differentiation of Distal Lung Epithelium from Embryonic Stem Cells
) w; k0 T8 o% vUNIT 1G.2 Pancreas Differentiation of Mouse ES Cells
# X" Q& q4 A2 f$ xUNIT 1G.3 Differentiation of Embryonic Stem Cells into Anterior Definitive Endoderm ) D/ G/ ], S* L, {: E5 F0 I$ x# V
Section H Ectodermal Lineages
. ^4 p5 Z5 |  ]  I- ?' tUNIT 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons
( m# u6 X. ^4 \4 _3 M( ?( G Chapter 2 Somatic Stem Cells + E8 W* m, j6 u; J/ B7 J- x
Section A Hematopoietic Stem Cells " [" Q, j- B* h( z  f
Hematopoietic Stem Cells
; m8 h/ O9 t  \6 i, |0 EUNIT 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll‐Paque Density Gradient - c0 Z, v; o/ `  o) @% N
UNIT 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood
4 N  W% O5 h- WUNIT 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood ; B5 G# W" w0 P1 j# ^9 f
UNIT 2A.4 Isolation and Assessment of Long‐Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow
; W7 g5 J1 l, e& M9 {; fUNIT 2A.5 Analysis of the Hematopoietic Stem Cell Niche
$ w# Z, E/ Y+ X$ LUNIT 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine Embryonic Tissues
" z7 Y- x; U( g% Q( z& OUNIT 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells
; Y1 [- Z% }3 _UNIT 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells   h" C! Q/ b) }$ H" F
Section B Non‐Hematopoietic Bone Marrow‐Derived Stem Cells : T0 z- d2 p( @7 e, r
UNIT 2B.1 Isolation and Characterization of Mesoangioblasts from Mouse, Dog, and Human Tissues ; I7 C9 I" h) @1 @  m3 r+ R4 r
UNIT 2B.2 Purification and Culture of Human Blood Vessel–Associated Progenitor Cells
% ~/ z& S) Y% W9 uUNIT 2B.3 Isolation, Culture, and Differentiation Potential of Mouse Marrow Stromal Cells ( P6 l# b0 R1 o9 E, r7 O  X0 z
Section C Cardiovascular Stem Cells
% r, L+ r% y5 ~/ l! w# _UNIT 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood ' W2 F" U. F5 I. p1 h* L: F3 ?
UNIT 2C.2 Derivation of Epicardium‐Derived Progenitor Cells (EPDCs) from Adult Epicardium + P4 X$ V' U: y9 v+ i0 V/ N
Section D Neural Stem Cells
# i# p/ f0 i% Q/ w4 q, QUNIT 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains
, Y) I+ Y/ N: t9 `UNIT 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells
# P3 t$ ]: c, P$ A" p/ o5 bUNIT 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 2 C/ l% v! X+ b! |  x  Y
Chapter 3 Cancer Stem Cells " ?$ I$ |) A& y, k% s6 }
UNIT 3.1 Colon Cancer Stem Cells
3 N) s& S2 m% k- z7 X  Y9 k% }. |UNIT 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model
* K2 W8 G7 B4 C1 p6 ` Chapter 4 Manipulation of Potency " H4 t) z2 K4 S2 F6 [1 l
Section A iPS Cells
0 c( M) w9 g, ~7 q8 ^/ ^UNIT 4A.1 Human iPS Cell Derivation/Reprogramming " J) q3 H& P+ m. E
UNIT 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells
7 L$ f( H9 c4 m; o Section B Nuclear Transfer
* ]: k5 y, B' `6 X. q+ d. eUNIT 4B.1 Heterokaryon‐Based Reprogramming for Pluripotency : Y% i& _0 C+ Z5 t
Chapter 5 Genetic Manipulation of Stem Cells 3 J6 }7 D5 [6 K! p7 r/ w; }
Section A Lineage Tracers in Stem Cells
& t. [; a# W. {' m1 K+ BUNIT 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma % G3 s. ?+ A6 c/ N8 d+ g; w
UNIT 5A.2 Functional Analysis of Adult Stem Cells Using Cre‐Mediated Lineage Tracing
. z# ~8 N9 u' Y8 f$ D1 y- u7 s$ y Appendix 1 Useful Information * }3 L: [! f  a1 T7 P5 b! {0 {, G
APPENDIX 1A Guidelines for the Conduct of Human Embryonic Stem Cell Research $ r0 g. x! z2 q5 Y; x; H
APPENDIX 1B ISSCR Guidelines for the Clinical Translation of Stem Cells # g6 ?9 p. ?. }7 w  b
Appendix 2 Laboratory Equipment ' J9 A0 ^, p: i
APPENDIX 2 Standard Laboratory Equipment
" a' j# [- X: P5 o$ a* s, o* ] Appendix Suppliers

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我有UNIT 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures