求助current protocols in stem cell biology

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Current Protocols in Stem Cell Biology. K/ n# J' U! H' h2 V% b7 w4 Q
Last Update: July 07, 2009/ J& ?1 [) ?. }+ x9 P/ @. h: C5 b
Print ISSN: 1941-7322( c& W  c' g7 b1 J% j/ Z
Online ISSN: 1938-8969
4 w! c- W" t, r3 x5 C& u4 c0 STable of Contents; c+ }% t  J5 F: Y% h5 E# p
Foreword
* N4 Q# q: J- P- `% E! O! M4 WPreface : e/ P; C* k: M( K) s' p( h9 Z
Chapter 1 Embryonic and Extraembryonic Stem Cells
  ^" g9 Y* m0 u$ ~0 ^) `- t; ? Section A Isolation of Embryonic Stem Cells . q" A) L) j1 S8 b- H
Isolation of Embryonic Stem Cells * y# T4 e8 u8 O. _" F) w5 Y
UNIT 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells
4 k+ T. h2 v3 M9 g! I. [' q; XUNIT 1A.2 Derivation of hESC from Intact Blastocysts 3 d9 D. Q2 _! w* |  O: z, U' o* q" D0 @
UNIT 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells
; A8 f6 `% e6 C$ I8 o& t5 NUNIT 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment
7 n  T; r4 v' F' A Section B Characterization of Embryonic Stem Cells
; u+ {8 z- M- h8 Y, [7 _: SUNIT 1B.1 Proteomic Analysis of Pluripotent Stem Cells 6 d; @. W- U: U- ~
UNIT 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and Embryoid Body–Derived Cells Using a High‐Throughput Microarray Platform
( d) p* a! B& X9 NUNIT 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells 3 k0 ]1 J; x2 H* F* F
UNIT 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency # y" J1 t1 _8 `& s
UNIT 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation ; p8 Y) `3 K3 i
Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells
  e5 Q% e& ?" b) tCulture and Maintenance of Undifferentiated Embryonic Stem Cells
0 `/ P6 o9 i' L! J3 wUNIT 1C.1 Expansion of Human Embryonic Stem Cells In Vitro
2 V) I9 \! L' V1 {/ mUNIT 1C.2 Defined, Feeder‐Independent Medium for Human Embryonic Stem Cell Culture
: w) A+ G+ E* G: p. x/ }' OUNIT 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells * R- I) Q7 j; _5 t
UNIT 1C.4 Culture of Mouse Embryonic Stem Cells
4 ~: V: g, d4 O6 D) hUNIT 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells
1 O0 @6 V- ^2 v8 d0 TUNIT 1C.6 Isolation of Human Placental Fibroblasts
, ?' U2 q3 P& s# x0 JUNIT 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells
; m% T9 `1 U0 O5 }4 X5 s  K& IUNIT 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of ROCK Inhibitor
5 n2 c/ O6 n) o, ] Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells
! N1 [1 O9 w% w. {6 N* g1 v* ]UNIT 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap 9 b. n. K& I1 h  h- c5 \- R
UNIT 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures
, X# U9 c  T+ O( Q$ B" J7 X* iUNIT 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay 6 q" `& c/ r; e3 i+ V* P% {* l
UNIT 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically Defined Culture Conditions 8 N& F4 L+ f" s' @* G  m
UNIT 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells $ H4 X0 U, _8 [( J8 E
Section E Extraembryonic Lineages
9 V$ j: Z% p  F4 T+ {/ ?" f- I6 I0 VIsolation of Stem Cells from Extraembryonic Tissues . I9 b3 Y# k& L) a6 n! ^
UNIT 1E.1 Isolation of Human Placenta‐Derived Multipotent Cells and In Vitro Differentiation into Hepatocyte‐Like Cells , E4 Q6 E7 Y5 C* r& L
UNIT 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta % K0 M, o0 h! _- p
UNIT 1E.3 Isolation of Amniotic Epithelial Stem Cells
/ v+ ?1 i, H7 a# @  [+ L4 g6 RUNIT 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells
& _% i# d! n- p- g Section F Mesodermal Lineages
4 G; i0 ~6 k$ B9 D6 S  e: ~UNIT 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells
. z" I- }5 J" f  Y) m4 q+ {0 uUNIT 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture with Endoderm in Serum‐Free Medium
9 M* y9 C0 W, h& a8 JUNIT 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells
+ k+ P" R% W8 D1 B9 o6 KUNIT 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood ' }( X  j, M2 Q+ H5 a) d
UNIT 1F.5 Endothelial Differentiation of Embryonic Stem Cells 8 Z5 o% \# I) n' q8 }) ~8 @( a, W
UNIT 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation with Stromal Layers
9 J* {( _' e8 _UNIT 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors
9 y3 j3 ]; y7 P" k& b" R5 lUNIT 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from Human Embryonic Stem Cells
0 C/ R; n& \! h Section G Endodermal Lineages
, a2 y  Z* A4 F' kUNIT 1G.1 The Differentiation of Distal Lung Epithelium from Embryonic Stem Cells
* X; K" E. c1 }& |' ~0 fUNIT 1G.2 Pancreas Differentiation of Mouse ES Cells # o) u) y. ~3 q" i  r% H
UNIT 1G.3 Differentiation of Embryonic Stem Cells into Anterior Definitive Endoderm
4 e8 `/ m9 q9 c5 t7 m/ J Section H Ectodermal Lineages 5 q4 B' T' \( p' i  G9 P2 r
UNIT 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons ' P9 z3 s( `. u% r
Chapter 2 Somatic Stem Cells - r! i3 ~. Z/ z8 o8 e
Section A Hematopoietic Stem Cells   O0 E+ g% m4 N% R# [& H( V
Hematopoietic Stem Cells
1 t# E8 i: O/ r2 X& g& xUNIT 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll‐Paque Density Gradient
( |( r" E) o/ s- u6 kUNIT 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood 1 Z4 D. f- ?- U$ `9 d
UNIT 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood & P; S/ g: q8 z$ s% G# m7 ^5 ^& c
UNIT 2A.4 Isolation and Assessment of Long‐Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow ) D3 u- W. [8 ?: b& n" u9 m
UNIT 2A.5 Analysis of the Hematopoietic Stem Cell Niche
) p* Q" [8 A- ~- jUNIT 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine Embryonic Tissues ' Y7 _4 i% }; R- L' N1 x2 P
UNIT 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells
" D8 a$ t' K; t7 K4 @0 K# q0 XUNIT 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells 8 {$ |* d( O# c- A0 m: U
Section B Non‐Hematopoietic Bone Marrow‐Derived Stem Cells
9 R: E2 M# E6 K- ^4 a* }( |UNIT 2B.1 Isolation and Characterization of Mesoangioblasts from Mouse, Dog, and Human Tissues
5 r( F4 U" }6 ^7 J/ P& m6 HUNIT 2B.2 Purification and Culture of Human Blood Vessel–Associated Progenitor Cells 1 {* ~" X9 T" o7 [
UNIT 2B.3 Isolation, Culture, and Differentiation Potential of Mouse Marrow Stromal Cells ; h& U8 d  c1 C' ], @8 G4 M. m5 ~) c
Section C Cardiovascular Stem Cells 4 [* j7 b) r" C; e8 [) ?7 Q: M7 V
UNIT 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood
$ j3 K" }# r% L. HUNIT 2C.2 Derivation of Epicardium‐Derived Progenitor Cells (EPDCs) from Adult Epicardium
& z1 p+ I6 m  ?9 p Section D Neural Stem Cells
1 B" H( N5 w2 tUNIT 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains 5 D! l+ W" q+ u9 ?6 p8 Z$ F
UNIT 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells 4 h3 k# m) Y' }! [  b
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 2 {6 z0 m4 @, l' y7 R5 i
Chapter 3 Cancer Stem Cells $ L" P# Q/ a9 Q
UNIT 3.1 Colon Cancer Stem Cells
: ~2 F$ C2 D3 x6 f# Q; MUNIT 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model
( i% ^$ B( L0 z! Q1 Q Chapter 4 Manipulation of Potency
- i2 u, ^7 \: ~, L Section A iPS Cells
! K3 n3 m+ U: rUNIT 4A.1 Human iPS Cell Derivation/Reprogramming
  Q1 p8 I9 @1 h4 cUNIT 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells 1 w+ D7 m( M4 J$ h" l, p7 y
Section B Nuclear Transfer
; I5 }% c/ E; |' bUNIT 4B.1 Heterokaryon‐Based Reprogramming for Pluripotency
7 {/ b5 e" f! O2 j! z Chapter 5 Genetic Manipulation of Stem Cells
5 l1 f. U9 U4 ~/ { Section A Lineage Tracers in Stem Cells
: z3 W3 R! E8 n( U( @8 w3 dUNIT 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma + M% D. P3 |& L( J& T. o2 N
UNIT 5A.2 Functional Analysis of Adult Stem Cells Using Cre‐Mediated Lineage Tracing
) z" S: C: i/ q$ l Appendix 1 Useful Information
2 r4 f6 x: \5 [: f' O! V  YAPPENDIX 1A Guidelines for the Conduct of Human Embryonic Stem Cell Research % ?& F8 f5 O, X1 ]  t
APPENDIX 1B ISSCR Guidelines for the Clinical Translation of Stem Cells
# x$ \2 f0 A* { Appendix 2 Laboratory Equipment
& `6 p7 J4 d) H+ N3 A5 F/ ^3 eAPPENDIX 2 Standard Laboratory Equipment
9 ?# s/ I& _  w* A. W  h Appendix Suppliers