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Current Protocols in Stem Cell Biology 2007年完整版 1451页: e6 ^& r" R  V$ s7 e/ Y
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Online ISBN:9780470151808
DOI: 10.1002/97804701518086 l% I/ d8 ?% A# B2 m
Table of Contents
1. Foreword7 I* S1 g, [- X- `) r2 Q; m
2. Preface7 a2 o7 e, `+ @7 j( G. W  z  v
3. Chapter 1 Embryonic and Extraembryonic Stem Cells
1. Section A Isolation of Embryonic Stem Cells
1. Introduction% i% P! Q& r% {) a
2. Unit 1A.1 Derivation and Characterization of Nonhuman Primate Embryonic Stem Cells6 v( n7 Y& n" R2 Y# t( }
3. Unit 1A.2 Derivation of hESC from Intact Blastocysts# a" E8 q( ^7 K1 ~5 @
4. Unit 1A.3 Reprogramming Primordial Germ Cells (PGC) to Embryonic Germ (EG) Cells
5. Unit 1A.4 Derivation and Propagation of hESC Under a Therapeutic Environment- K  w. t# p$ m$ N8 D3 |9 {( \; _
2. Section B Characterization of Embryonic Stem Cells) f$ i4 K' u* Z: Z5 ~7 l4 e' Y
1. Unit 1B.1 Proteomic Analysis of Pluripotent Stem Cells
2. Unit 1B.2 Gene Expression Analysis of RNA Purified from Embryonic Stem Cells and
Embryoid Body–Derived Cells Using a High-Throughput Microarray Platform
3. Unit 1B.3 Phenotypic Analysis of Human Embryonic Stem Cells
4. Unit 1B.4 Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of: N1 W' A+ G* ?
Pluripotency2 W) i0 k1 P3 n# k7 Y( q
5. Unit 1B.5 Tandem Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells
Using In Vivo Biotinylation
6. Unit 1B.6 Characterization of X-Chromosome Inactivation Status in Human Pluripotent Stem
Cells
7. Unit 1B.7 Preparation of Defined Human Embryonic Stem Cell Populations for Transcriptional* V# l) M) j2 r% g* y9 N
Profiling
3. Section C Culture and Maintenance of Undifferentiated Embryonic Stem Cells
1. Introduction
2. Unit 1C.1 Expansion of Human Embryonic Stem Cells In Vitro
3. Unit 1C.2 Defined, Feeder-Independent Medium for Human Embryonic Stem Cell Culture" t4 ^" t4 |8 @0 P
4. Unit 1C.3 Isolation and Propagation of Mouse Embryonic Fibroblasts and Preparation of' ^7 O. m9 K; ?& E$ W2 T  X
Mouse Embryonic Feeder Layer Cells
5. Unit 1C.4 Culture of Mouse Embryonic Stem Cells
6. Unit 1C.5 Preparation of Autogenic Human Feeder Cells for Growth of Human Embryonic, z% o0 a$ z" f6 g0 O, _: j0 Q$ ^
Stem Cells
7. Unit 1C.6 Isolation of Human Placental Fibroblasts
8. Unit 1C.7 Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic6 g4 v% X9 A2 D5 n
Stem Cells
9. Unit 1C.8 Cryopreservation of Dissociated Human Embryonic Stem Cells in the Presence of5 I* G1 K) H# Y: G% d2 n9 i
ROCK Inhibitor( ?; u# R. C4 [7 o+ y) T
10. Unit 1C.9 Authentication and Banking of Human Pluripotent Stem Cells: \& E3 A, E4 S. ^2 v
11. Unit 1C.10 Clump Passaging and Expansion of Human Embryonic and Induced Pluripotent
Stem Cells on Mouse Embryonic Fibroblast Feeder Cells/ @/ [! P3 f2 h1 [
12. Unit 1C.11 Expansion of Human Embryonic Stem Cells on Cellulose Microcarriers9 v& W% H+ q- D
4. Section D Germ Layer Induction/Differentiation of Embryonic Stem Cells- ]6 G9 y# Y* p+ q& c/ L
1. Unit 1D.1 Germ Layer Induction in ESC—Following the Vertebrate Roadmap
2. Unit 1D.2 Formation and Hematopoietic Differentiation of Human Embryoid Bodies by
Suspension and Hanging Drop Cultures/ ]+ B! k5 u) N  v( |
3. Unit 1D.3 Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies! M4 i" V8 f3 c7 z% O7 V# k0 Y
and a Description of the Hematopoietic Blast Colony Forming Assay
4. Unit 1D.4 Differentiation of Human Embryonic Stem Cells in Adherent and in Chemically
Defined Culture Conditions- y7 A! _( d2 s# B% g4 N
5. Unit 1D.5 Isolation and Differentiation of Xenopus Animal Cap Cells
5. Section E Extraembryonic Lineages
1. Introduction
2. Unit 1E.1 Isolation of Human Placenta-Derived Multipotent Cells and In Vitro Differentiation+ {3 a0 ^# w$ w$ S  i( j$ F% Q3 O+ p
into Hepatocyte-Like Cells
3. Unit 1E.2 Isolation of Mesenchymal Stem Cells from Amniotic Fluid and Placenta
4. Unit 1E.3 Isolation of Amniotic Epithelial Stem Cells: u& [: V5 p! n' p- F, z2 Q0 b
5. Unit 1E.4 Isolation and Manipulation of Mouse Trophoblast Stem Cells$ W4 |! P  L8 N0 c
6. Unit 1E.5 Isolation of Amniotic Mesenchymal Stem Cells
7. Unit 1E.6 Amnion Epithelial Cell Isolation and Characterization for Clinical Use
6. Section F Mesodermal Lineages: E, X/ g+ d  T+ N8 m
1. Unit 1F.1 Differentiation of Embryonic Stem Cells into Cartilage Cells
2. Unit 1F.2 Differentiation of Human Embryonic Stem Cells to Cardiomyocytes by Coculture, z4 H' `9 g8 n0 F* r0 e
with Endoderm in Serum-Free Medium
3. Unit 1F.3 Isolation of Hematopoietic Stem Cells from Mouse Embryonic Stem Cells# p8 s5 P! L/ Q" ]3 f8 z. l; N
4. Unit 1F.4 Differentiation of Mouse Embryonic Stem Cells into Blood
5. Unit 1F.5 Endothelial Differentiation of Embryonic Stem Cells
6. Unit 1F.6 Hematopoietic Differentiation of Human Embryonic Stem Cells by Cocultivation- J( b- v- ^  N/ R: g* Y7 Q7 y7 v
with Stromal Layers
7. Unit 1F.7 TLX1 (HOX11) Immortalization of Embryonic Stem Cell–Derived and Primary9 s# |9 c9 Z- X/ _# y/ W$ y
Murine Hematopoietic Progenitors
8. Unit 1F.8 Differentiation of Multipotent Mesenchymal Precursors and Skeletal Myoblasts from( E: _  r* ]4 D9 u6 x, ?: W
Human Embryonic Stem Cells
9. Unit 1F.9 Derivation of Vasculature from Embryonic Stem Cells
10. Unit 1F.10 Isolation and Functional Characterization of Pluripotent Stem Cell–Derived Cardiac6 Y, o' d. f9 P1 r0 K
Progenitor Cells5 G5 a" e$ o3 N& `2 O
11. Unit 1F.11 Differentiation of Mouse Embryonic Stem Cells into Cardiomyocytes via the; U- {. b, Y8 k. O$ h
Hanging-Drop and Mass Culture Methods
7. Section G Endodermal Lineages
1. Unit 1G.1 The Differentiation of Distal Lung Epithelium from Embryonic Stem Cells
2. Unit 1G.2 Pancreas Differentiation of Mouse ES Cells
3. Unit 1G.3 Differentiation of Embryonic Stem Cells into Anterior Definitive Endoderm6 g% V% O  g( A' k2 P, J
8. Section H Ectodermal Lineages1 x1 L8 m2 t! G3 Z& w
1. Unit 1H.1 Differentiation of Mouse Embryonic Stem Cells to Spinal Motor Neurons" P$ w5 g2 B* S7 t$ g) s  M5 m
2. Unit 1H.2 Time-Lapse Imaging of Embryonic Neural Stem Cell Division in Drosophila by) d* R/ f, _0 ^- {
Two-Photon Microscopy
4. Chapter 2 Somatic Stem Cells
1. Section A Hematopoietic Stem Cells
1. Introduction2 M9 S* q& T* X
2. Unit 2A.1 Isolation of Mononuclear Cells from Human Cord Blood by Ficoll-Paque Density) t. S) K1 f  |. u0 q9 O
Gradient: i8 Y" A, p2 G9 D9 v
3. Unit 2A.2 Isolation of Hematopoietic Stem Cells from Human Cord Blood
4. Unit 2A.3 Isolation of Mesenchymal Stem Cells from Human Cord Blood
5. Unit 2A.4 Isolation and Assessment of Long-Term Reconstituting Hematopoietic Stem Cells, L1 M2 Y! `$ x7 X/ G
from Adult Mouse Bone Marrow# L5 $ J! x6 @9 C
6. Unit 2A.5 Analysis of the Hematopoietic Stem Cell Niche
7. Unit 2A.6 Analysis and Manipulation of Hematopoietic Progenitor and Stem Cells from Murine8 h+ b4 Q. M2 N
Embryonic Tissues6 U/ t9 g# G$ A* ^- n% p
8. Unit 2A.7 High Level In Vitro Expansion of Murine Hematopoietic Stem Cells) `' Z+ }, o2 \$ H( W1 o
9. Unit 2A.8 Isolation and Visualization of Mouse Placental Hematopoietic Stem Cells, $ A1 ^( j8 P- J2 v/ c. j
10. Unit 2A.9 Preparation of Hematopoietic Stem and Progenitor Cells from the Human Placenta
2. Section B Non-Hematopoietic Bone Marrow-Derived Stem Cells
1. Unit 2B.1 Isolation and Characterization of Mesoangioblasts from Mouse, Dog, and Human. W9 }" [5 c& X4 N9 r( I
Tissues
2. Unit 2B.2 Purification and Culture of Human Blood Vessel–Associated Progenitor Cells
3. Unit 2B.3 Isolation, Culture, and Differentiation Potential of Mouse Marrow Stromal Cells0 l! ^6 c$ d$ {4 p1 r
3. Section C Cardiovascular Stem Cells' \1 M# Y2 d* b7 o; T
1. Unit 2C.1 Isolation and Characterization of Endothelial Progenitor Cells from Human Blood. K. I; l  o) R7 {
2. Unit 2C.2 Derivation of Epicardium-Derived Progenitor Cells (EPDCs) from Adult Epicardium, L3 [" C; B. f  s+ y6 B4 e3 V
3. Unit 2C.3 Isolation and Expansion of Cardiosphere-Derived Stem Cells7 [* C" S, K) ]6 a
4. Section D Neural Stem Cells
1. Unit 2D.1 Generation of Cultured Oligodendrocyte Progenitor Cells from Rat Neonatal Brains- f: {) s9 N7 m  ^) @
2. Unit 2D.2 Isolating, Expanding, and Infecting Human and Rodent Fetal Neural Progenitor Cells5 a- x! ^3 E- _; _) d
3. Unit 2D.3 Long-Term Multilayer Adherent Network (MAN) Expansion, Maintenance, and: E2 D7 Z8 h, J% v; r
Characterization, Chemical and Genetic Manipulation, and Transplantation of Human Fetal) w2 `# I( V2 n. z1 o. A' d
Forebrain Neural Stem Cells3 z" M# {: n/ c
4. Unit 2D.4 Culture System for Rodent and Human Oligodendrocyte Specification, Lineage
Progression, and Maturation$ j: n5 k$ ^2 e) v2 z' r
5. Unit 2D.5 Isolation and Culture of Ventral Mesencephalic Precursor Cells and Dopaminergic
Neurons from Rodent Brains- ]# j, n! K  N
6. Unit 2D.6 Isolation of Neural Stem Cells from Neural Tissues Using the Neurosphere
Technique
5. Section E Germline Stem Cells
1. Unit 2E.1 Culturing Ovarian Somatic and Germline Stem Cells of Drosophila- s7 h- H" j- w$ @6 M& b
2. Unit 2E.2 Time-Lapse Live Imaging of Stem Cells in Drosophila Testis* B; j& U# T  m2 {* Z# J/ l0 h
6. Section F Gut Stem Cells# s- [& q  V9 j
1. Unit 2F.1 In Situ Hybridization to Identify Gut Stem Cells( Z2 u, I! h1 X+ v
7. Section G Lung Stem Cells
1. Unit 2G.1 Isolation and Clonal Assay of Adult Lung Epithelial Stem/Progenitor Cells
5. Chapter 3 Cancer Stem Cells
1. Unit 3.1 Colon Cancer Stem Cells
2. Unit 3.2 In Vivo Evaluation of Leukemic Stem Cells through the Xenotransplantation Model, j/ x8 z" c" q! D
3. Unit 3.3 Culture and Isolation of Brain Tumor Initiating Cells
6. Chapter 4 Manipulation of Potency
1. Section A iPS Cells) d& r$ |( E9 F7 J# v0 `, m8 E
1. Unit 4A.1 Human iPS Cell Derivation/Reprogramming7 c9 X7 e! Q5 G
2. Unit 4A.2 Generation and Characterization of Human Induced Pluripotent Stem Cells
2. Section B Nuclear Transfer
1. Unit 4B.1 Heterokaryon-Based Reprogramming for Pluripotency. ]0 h* b9 O" L5 t: C6 m& }
7. Chapter 5 Genetic Manipulation of Stem Cells
1. Section A Lineage Tracers in Stem Cells' k% [2 r0 Z3 `& L5 C% T
1. Unit 5A.1 Imaging Neural Stem Cell Fate in Mouse Model of Glioma" b, Z( t& G9 x" X: Q) O. L
2. Unit 5A.2 Functional Analysis of Adult Stem Cells Using Cre-Mediated Lineage Tracing1 v+ d4 K0 W9 u+ w3 Q
3. Unit 5A.3 Magnetic Resonance Imaging of Human Embryonic Stem Cells1 u: j! h; {: Z! K
4. Unit 5A.4 Lineage Tracing in the Intestinal Epithelium
5. Unit 5A.5 Kaede-Centrin1 Labeling of Mother and Daughter Centrosomes in Mammalian' {+ s" ^) p, M  m
Neocortical Neural Progenitors
2. Section B Homologous Recombination in Stem Cells
6. Unit 5B.1 Generation of Human Embryonic Stem Cell Reporter Knock-In Lines by
Homologous Recombination
8. Appendix 1 Useful Information  y6 y$ o, I7 ^) f/ p' c
1. 1A Guidelines for the Conduct of Human Embryonic Stem Cell Research( |- w$ V$ j3 o+ U$ c% I
2. 1B ISSCR Guidelines for the Clinical Translation of Stem Cells  l4 f5 J# B4 n% E6 Y
9. Appendix 2 Laboratory Equipment/ ^5 N5 U+ O0 v% l
Standard Laboratory Equipment
10. Appendix Suppliers) ]. O& j/ O& Y# Z* f3 ]* W
Selected Suppliers of Reagents and Equipment. f+ }; b. h- c$ g' ]

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