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发表于 2012-1-13 13:55 |只看该作者 |倒序浏览 |打印

本帖最后由细胞海洋于 2012-1-13 14:38 编辑 / x2 N0 W% t/ t. s. _

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Springer出版社的新书，很多图，对做人多能干细胞的童鞋可能有用。
ISBN 978-1-61779-547-3 e-ISBN 978-1-61779-548-0# U1 x9 w% O0 s
DOI 10.1007/978-1-61779-548-0
Springer New York Dordrecht Heidelberg London, e0 J5 |# z/ c! S6 o
Library of Congress Control Number: 20119416082 Q$ F9 x$ @9 W' u* v7 z  r
© Springer Science+Business Media, LLC 2012
[hide][/hide]! I! P. {8 ^4 \# @  x
Contents：
1Methods for the Derivation of Human Embryonic$ k1 K% W# L- V; x2 L4 b. g
Stem Cell Lines .......................................................................................... 1& M4 U1 i5 f, V$ L+ ^3 _
1.1 Introduction ........................................................................................ 1
1.2 Materials for ESC Line Derivation .................................................... 9
1.3 Methods for hESC Isolation .............................................................. 9% ?% D' q7 V$ V3 q0 o
1.3.1 hESC Isolation by Immunosurgery........................................ 125 k' M0 Q+ `7 F( g( J. O
1.3.2 Mechanical Removal of Trophectoderm ................................ 12- A+ K! \/ u6 c5 u8 O, }& C
1.3.3 Whole Embryo Approach for ESC Line Derivation .............. 13
References ................................................................................................... 13
2 Morphology of Human Embryonic and Induced Pluripotent
Stem Cell Colonies Cultured with Feeders ............................................. 15+ n' `+ D) @" H! j
2.1 Introduction ........................................................................................ 15
2.2 Materials ............................................................................................ 16
2.2.1 For Mouse Embryonic Fibroblasts (MEFs)+ [- [" S$ B3 u
and Foreskin Fibroblasts (HFFs) ........................................... 16
2.2.2 For hPSC Maintenance .......................................................... 17/ R8 N4 E# y2 q
2.3 Methods ............................................................................................. 185 R, k# X: M2 [8 h+ z8 p: o: v7 c
2.3.1 Feeder Culture Methods ........................................................ 18' h. m" E' }( J* H9 E
2.3.2 hPSC Culture ......................................................................... 223 {9 M2 w5 E2 x+ P+ {
References ................................................................................................... 38
3 Morphology of Human Embryonic Stem Cells and Induced: T/ r6 c" s) e9 K( w
Pluripotent Stem Cells Cultured in Feeder8 @1 e4 j/ Z& R9 K4 B7 ?
Layer-Free Conditions .............................................................................. 41
3.1 Introduction ........................................................................................ 41
3.2 Materials for Feeder Layer-Free Culture of hPSCs ........................... 43
3.2.1 Matrix Preparation ................................................................. 43
3.2.2 Culture Medium ..................................................................... 44
3.3 Methods for hPSC Feeder Layer-Free Culture .................................. 44
3.3.1 Preparation of Matrix-Covered Plates ................................... 44& ~/ A4 u$ ?5 v$ }: i* ^$ |! R0 ?% `
3.3.2 Splitting, Freezing, and Thawing hPSCs ............................... 45
3.3.3 Adaptation of PSCs to Feeder-Free Culture .......................... 45. O  a, ^6 A8 [" G1 L. y7 N! O
3.3.4 Routine Culture of hPSCs ...................................................... 46
References ................................................................................................... 540 V; Z: }* t" D+ _
4 Morphology of Undifferentiated Human Embryonic% C+ @6 j2 I" @. L5 a  Q
and Induced Stem Cells Grown in Suspension
and in Dynamic Cultures .......................................................................... 57
4.1 Introduction ........................................................................................ 57
4.2 Materials for Suspension Culture of hPSCs ...................................... 587 a) d: [, _% ?% b# C
4.2.1 Culture Medium ..................................................................... 58, ]) h3 o; q) U$ b/ N4 k
4.2.2 Splitting Medium ................................................................... 59
4.2.3 Freezing Medium ................................................................... 59
4.3 Methods for Suspension Culture of hPSCs ....................................... 605 w9 F/ e' n+ B; g1 ?! R3 a
4.3.1 Creating a hPSC Suspension Culture .................................... 60% m$ N7 G4 c- l0 `) J
4.3.2 Splitting hPSCs in Suspension ............................................... 60! e3 L3 M# Y( Y) @+ J
4.3.3 Freezing hPSCs in Suspension .............................................. 62
4.3.4 Thawing hPSCs in Suspension .............................................. 646 d6 i: A8 j% d' l1 Q( }
4.3.5 Culturing hPSCs in a Dynamic System ................................. 64
4.3.6 Routine Culture of hPSCs in Suspension .............................. 65
References ................................................................................................... 71. u4 E# E: [1 C" j; B
5 Differentiation of Pluripotent Stem Cells In Vitro:3 ~7 z5 v8 F# t# G2 O- M! J
Embryoid Bodies ....................................................................................... 736 G2 j: r2 [! l1 B
5.1 Introduction ........................................................................................ 73
5.2 Materials for EB Formation ............................................................... 750 u6 F5 n* I' L* p8 o2 v
5.2.1 Culture Medium Supplemented with Serum ......................... 75
5.2.2 Splitting Medium Based on Collagenase ............................... 764 q% A5 D( t2 t- I7 \; s1 a
5.3 Methods for EB Formation and Culture ............................................ 76
5.3.1 EB Formation ......................................................................... 76
5.3.2 Routine Culture of EBs .......................................................... 768 g; `8 X9 M3 V8 |, S& V
5.3.3 Culturing EBs in Spinner Flasks ............................................ 787 T0 {; b2 W) k7 L6 i5 V, W0 R
References ................................................................................................... 88
6 Differentiation of Pluripotent Stem Cells In Vivo:
Teratoma Formation ................................................................................. 919 s3 x, Q7 O! A# r6 |
6.1 Introduction ........................................................................................ 91
6.2 Materials for Teratoma Formation ..................................................... 93
6.2.1 Culture Medium ..................................................................... 930 S6 [( H9 K+ F
6.2.2 Syringe for Injecting Cells ..................................................... 93
6.3 Formation of Teratomas ..................................................................... 933 T$ G  E5 l2 o  I' H1 K% L) Q
6.3.1 Protocol for Teratoma Formation .......................................... 93. _3 H9 K0 \% n' W+ c9 K2 c
6.3.2 Routine Treatment of Mice and Teratoma ............................. 930 G1 |$ _9 r9 X! Q5 G" k
References ................................................................................................... 103( d- K1 M, K; q  C9 u! r
7 Immunostaining ........................................................................................ 105  l, F, H, t- j& R* X
7.1 Introduction ........................................................................................ 105% U8 P7 G* K" J# l5 _8 t& P
7.2 Materials and Solutions for Immunostaining .................................... 111
7.2.1 Materials and Solutions for Immunohistochemistry
of Paraffi n-Embedded Tissues ............................................... 111" F, K, g0 R1 h8 m* J2 h1 o
7.2.2 Materials and Solutions for Immunofl uorescence ................. 1114 {9 G! \1 \' }- i. Q' c' {
7.3 Immunostaining Procedures .............................................................. 112
7.3.1 Immunohistochemistry of Paraffi n-Embedded Tissues ......... 112
7.3.2 Immunofl uorescence of Cultured Cells ................................. 113
References ................................................................................................... 113- K+ d" N+ R: d6 m
8 Karyotype and Fluorescent In Situ Hybridization, l) L3 n/ [, N, O
Analysis of Human Embryonic Stem Cell and Induced
Pluripotent Stem Cell Lines ..................................................................... 115- J1 i+ E( \6 n. W
8.1 Introduction ........................................................................................ 1157 K7 |) s2 ~: @. W, s
8.1.1 Karyotype Analysis ............................................................... 1152 Q* M* t- v8 n' c! A1 V
8.1.2 FISH Analysis ........................................................................ 121+ f0 K+ P3 K1 y$ `/ d
8.2 Materials for Harvesting Cells for Karyotyping
and FISH Analysis ............................................................................. 124
8.2.1 Reagents ................................................................................. 1249 N4 w( F5 C# ^; z1 U
8.2.2 Solutions ................................................................................ 124
8.3 Procedure of Harvesting Cells for Karyotyping9 ?6 F  R% M+ h) ]/ f
and FISH Analysis ............................................................................. 124) ~+ \; K! V( U
References ................................................................................................... 126
9 Method for the Derivation of Induced Pluripotent5 D) Y8 {# W. T) h
Stem Cells from Human Hair Follicle Keratinocytes ............................ 127
9.1 Introduction ........................................................................................ 127- M# _8 z3 `2 W* ^! o5 ]) V7 [* U
9.2 Materials ............................................................................................ 1294 }5 W3 ]. b$ u! F# Z# v
9.2.1 NIH-3T3/293T Cells .............................................................. 129
9.2.2 Keratinocyte Derivation from Plucked Hair Follicles ........... 129
9.3 Methods ............................................................................................. 130
9.3.1 NIH-3T3 and 293T Culture Methods .................................... 130) Y' `- F; X; d) w
9.3.2 Keratinocyte Culture Methods ............................................... 1329 q1 n/ _- ]; \# ~1 ?
9.3.3 Preparation of the STEMCCA Virus for Infection ................ 133' l8 Q7 \+ `; L, s8 }9 l
9.3.4 Derivation of iPSCs from Hair Keratinocytes ....................... 1342 a( S  e6 E/ v5 n* o. a6 D( ?7 O
References ................................................................................................... 137

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