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5 _1 b; n" P8 @" F% B, E# `( C
! N. {0 ~ K+ o {; dCurrent Protocols in Stem Cell Biology
/ q+ F9 a) H4 J: d% VLast Update: July 07, 20090 j4 B; Y5 z; L" P
Print ISSN: 1941-7322% ]& ? S6 t) B) S& n$ E! E
Online ISSN: 1938-8969( I! ]( l) b! _$ f1 j! H, \' o
Table of Contents
: w% d3 j7 j. ~# TForeword
! B3 `3 _ P9 SPreface & D- S/ D. l3 |
Chapter 1 Embryonic and Extraembryonic Stem Cells
: K+ K% q0 P7 c8 A p+ N Section A Isolation of Embryonic Stem Cells
; a6 s" Y( ~1 t# A* D4 X* w8 `7 {. QIsolation of Embryonic Stem Cells
: R* O$ h% ~0 C: \UNIT 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells
& R9 P8 e% X3 v6 R, K! s3 EUNIT 1A.2 Derivation of hESC from Intact Blastocysts
- G- J3 H" R' u% FUNIT 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells 1 J6 q8 J' }6 t" s
UNIT 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment 9 ]! d2 B( \& Y6 A" P
Section B Characterization of Embryonic Stem Cells % i$ N/ Q8 ^/ g1 g5 m5 t7 c
UNIT 1B.1 Proteomic Analysis of Pluripotent Stem Cells & ?' _( s! `9 E: y7 G
UNIT 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and Embryoid Body–Derived Cells Using a High‐Throughput Microarray Platform a$ V! P7 i. N* r H# E
UNIT 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells 3 X( k4 l4 V3 o7 h& t& T
UNIT 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency 5 S9 i4 F6 D! z" z# r
UNIT 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells Using In Vivo Biotinylation
% Q9 O+ u0 r0 i0 K2 {3 a5 Y+ i Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells % R, D; C: D! K$ k
Culture and Maintenance of Undifferentiated Embryonic Stem Cells
# ^. ^% u% |& gUNIT 1C.1 Expansion of Human Embryonic Stem Cells In Vitro ! c, V l! j0 A- t* |, \+ U
UNIT 1C.2 Defined, Feeder‐Independent Medium for Human Embryonic Stem Cell Culture ' A7 X& f) L1 C
UNIT 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of Mouse Embryonic Feeder Layer Cells
( g. q, ?: ~4 o+ J! N, `5 P0 {UNIT 1C.4 Culture of Mouse Embryonic Stem Cells
* f& [! b) q6 u& i, A5 G) |: vUNIT 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic Stem Cells
$ o; y: e W( W8 `UNIT 1C.6 Isolation of Human Placental Fibroblasts
& u% ~+ f/ U+ ?) wUNIT 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells , q' d" `8 f2 M) A2 P; j
UNIT 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of ROCK Inhibitor
8 j/ U9 D. _- U: ~4 L) U Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells
8 `( }9 h F; `5 d4 ^UNIT 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap
5 F8 [4 r0 D2 @0 K/ j# H! t1 WUNIT 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by Suspension and Hanging Drop Cultures ! B; x$ N- @" k! E$ C) z. H, Y
UNIT 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay % H. r& V4 h6 R) B. f
UNIT 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically Defined Culture Conditions ! R* J7 F C" J7 x- S5 i
UNIT 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells 1 G4 |8 p" s z! X6 r2 C: l
Section E Extraembryonic Lineages 8 s0 T: n" T" x( f/ s
Isolation of Stem Cells from Extraembryonic Tissues
% U7 p0 N7 k$ a8 w( ~UNIT 1E.1 Isolation of Human Placenta‐Derived Multipotent Cells and In Vitro Differentiation into Hepatocyte‐Like Cells
" Q3 Q5 c! k% [. J; T% LUNIT 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta
+ `3 r& Y3 {: p" ?. g% G$ QUNIT 1E.3 Isolation of Amniotic Epithelial Stem Cells
* P" ?" ]" D( MUNIT 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells
& H Z: S: a' q# P9 |, [+ H Section F Mesodermal Lineages
0 F7 f9 l9 S" N6 n% b4 O* Q+ R0 U+ {UNIT 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells
# d7 j/ ~6 @ q5 Z3 JUNIT 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture with Endoderm in Serum‐Free Medium ! j2 n. C' B/ N1 @5 |/ s+ l6 N
UNIT 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells ( B% |& j8 g: |$ `( R
UNIT 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood
+ r9 N2 {; ^) C" p; m t6 c+ AUNIT 1F.5 Endothelial Differentiation of Embryonic Stem Cells
( g9 Q. j9 z+ K, B. a- DUNIT 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation with Stromal Layers 7 R' u2 d u, ~# A, a% m+ Y3 N
UNIT 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary Murine Hematopoietic Progenitors 1 m( c. }; D) Z4 S. y
UNIT 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from Human Embryonic Stem Cells 1 _) ?4 \6 I( `% ~$ e
Section G Endodermal Lineages ; j$ p0 w- d8 U9 Z' R
UNIT 1G.1 The Differentiation of Distal Lung Epithelium from Embryonic Stem Cells
P& N- D% Q% F2 [3 S7 u9 JUNIT 1G.2 Pancreas Differentiation of Mouse ES Cells
0 p' p9 M4 _0 y! VUNIT 1G.3 Differentiation of Embryonic Stem Cells into Anterior Definitive Endoderm ) e$ d) e7 _& }$ j: Z
Section H Ectodermal Lineages ) _8 l. ?' Z! W% X2 ]. r
UNIT 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons 1 m2 N6 U& } P
Chapter 2 Somatic Stem Cells
9 T* G6 Y! C4 D% F" H' J Section A Hematopoietic Stem Cells + I% x0 T5 @: W6 `5 q) W
Hematopoietic Stem Cells $ R0 {# J2 l7 W: L% ^% E
UNIT 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll‐Paque Density Gradient $ j3 U+ E( E7 s. M3 C7 g
UNIT 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood
2 I' L$ }* q X5 Z! mUNIT 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood
: o; K& h$ Y( F! t. ? _3 pUNIT 2A.4 Isolation and Assessment of Long‐Term Reconstituting Hematopoietic Stem Cells from Adult Mouse Bone Marrow
# G' F, w$ ?# I" w. |UNIT 2A.5 Analysis of the Hematopoietic Stem Cell Niche
* z5 g" _8 M8 I3 Y1 j) f4 d4 zUNIT 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine Embryonic Tissues 5 o2 U2 S$ a. `) }# o+ o7 ~2 n
UNIT 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells
2 _) ~3 V4 U4 F$ s% MUNIT 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells & y6 R; V3 O: M. M; T2 Q: d
Section B Non‐Hematopoietic Bone Marrow‐Derived Stem Cells
: P* {. e; |& D5 V0 UUNIT 2B.1 Isolation and Characterization of Mesoangioblasts from Mouse, Dog, and Human Tissues
& i$ T& J) E8 |0 [ [9 a& ~UNIT 2B.2 Purification and Culture of Human Blood Vessel–Associated Progenitor Cells
0 X' h/ y0 [6 Z# }UNIT 2B.3 Isolation, Culture, and Differentiation Potential of Mouse Marrow Stromal Cells * Q I, x; o2 Q& k4 ~( e3 h
Section C Cardiovascular Stem Cells 4 C- r+ v1 @6 h! }
UNIT 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood 8 |# }4 P: Y [4 \. `
UNIT 2C.2 Derivation of Epicardium‐Derived Progenitor Cells (EPDCs) from Adult Epicardium
* h a i, Z( ~2 p5 D, J P Section D Neural Stem Cells + Y6 {: m, n% w
UNIT 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains / k; K: w& i6 ^/ t9 R$ t: c3 b
UNIT 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells ; m4 j0 u# q5 |
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
* Q' Q) i: R# M/ i" q; x2 _ Chapter 3 Cancer Stem Cells 7 }; v8 C# n" q/ A1 a
UNIT 3.1 Colon Cancer Stem Cells 2 r R( R. r* X8 K B
UNIT 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model
" t% S- a% h. v# @. [1 N) }8 x Chapter 4 Manipulation of Potency
6 A/ w" B: N8 H; \( Z# {9 n Section A iPS Cells 1 j/ ]" l7 B, R6 Q2 N* c
UNIT 4A.1 Human iPS Cell Derivation/Reprogramming ' Z% b7 z+ Z$ B* w6 e, n
UNIT 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells
" t' E) X, E# K' v x# V" u Section B Nuclear Transfer
" C9 K4 w1 _7 z; H& V, j% m7 I4 oUNIT 4B.1 Heterokaryon‐Based Reprogramming for Pluripotency 9 ?7 @- R4 s' y% ?
Chapter 5 Genetic Manipulation of Stem Cells 9 ?6 ]# u/ r4 g+ E2 t+ }$ h
Section A Lineage Tracers in Stem Cells # j* R1 D1 {1 I: ~
UNIT 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma ; ]: E* m: |& g3 U5 J& Y
UNIT 5A.2 Functional Analysis of Adult Stem Cells Using Cre‐Mediated Lineage Tracing
; ^1 e- w& G9 f8 S8 V9 i Appendix 1 Useful Information
3 G' T( G. Q) h# R9 V& KAPPENDIX 1A Guidelines for the Conduct of Human Embryonic Stem Cell Research D- U3 J8 o+ x4 E+ v8 V: J& l
APPENDIX 1B ISSCR Guidelines for the Clinical Translation of Stem Cells
6 M9 V$ {6 H" a* I# s. c Appendix 2 Laboratory Equipment
, {3 P7 z: A' A z5 y9 Y7 eAPPENDIX 2 Standard Laboratory Equipment , \6 M2 W K+ Q2 C4 k: W
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发表于 2009-7-28 16:42
发表于 2009-7-29 16:23
