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本帖最后由 细胞海洋 于 2012-1-13 14:38 编辑 / C. U# q' J `1 I( ~# q6 U1 C5 W
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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。 o6 D; }) q4 y f ?7 S. ~4 F, [
ISBN 978-1-61779-547-3 e-ISBN 978-1-61779-548-0
2 h; E0 J9 X6 @/ e( FDOI 10.1007/978-1-61779-548-0
2 E; C Z4 c4 Z8 s' [Springer New York Dordrecht Heidelberg London
. G* g- m+ m6 zLibrary of Congress Control Number: 2011941608
) N' B1 c& V% u+ m© Springer Science+Business Media, LLC 2012
& g K7 |# N+ o- R) u* s+ {% `& ?6 [4 M[hide][/hide]
1 v: C0 c) x0 @# TContents：' v$ k7 J3 `4 R: n/ s
1Methods for the Derivation of Human Embryonic# x6 v) q. L; w$ }* q! k4 \
Stem Cell Lines .......................................................................................... 1. W: \5 y* P6 E. r6 ? b9 Q" d o2 c
1.1 Introduction ........................................................................................ 11 r/ X/ ~3 @% t0 A
1.2 Materials for ESC Line Derivation .................................................... 9
2 f, a2 H8 m$ e& D/ A1.3 Methods for hESC Isolation .............................................................. 9
; D7 \ ^) S' B# g: e: W1.3.1 hESC Isolation by Immunosurgery........................................ 120 Q' ?. P g5 I# T4 ^
1.3.2 Mechanical Removal of Trophectoderm ................................ 12' |2 `" n% I6 c3 ?
1.3.3 Whole Embryo Approach for ESC Line Derivation .............. 13( [+ a, e4 j0 p3 G- S; @
References ................................................................................................... 137 o R; t# X( }1 H+ }
2 Morphology of Human Embryonic and Induced Pluripotent
: _. n8 L" z- A+ y8 MStem Cell Colonies Cultured with Feeders ............................................. 15
+ R1 X% l# r! i! x7 g2.1 Introduction ........................................................................................ 15- L, l% Z& ?3 p" E+ ]6 r% A
2.2 Materials ............................................................................................ 16
}# S3 a4 k1 U2 j# N2.2.1 For Mouse Embryonic Fibroblasts (MEFs)
9 X. b S. ~3 q9 _& fand Foreskin Fibroblasts (HFFs) ........................................... 16* `& g8 y7 V- m- z! q; R
2.2.2 For hPSC Maintenance .......................................................... 177 W+ U7 q4 y7 |3 V' f8 y, q8 j
2.3 Methods ............................................................................................. 18
" M/ C# v0 O I/ a# H4 b2.3.1 Feeder Culture Methods ........................................................ 18
, N1 e8 m9 H% j2.3.2 hPSC Culture ......................................................................... 22
# t/ R1 Z8 ?; {: X o2 \4 ?, |2 WReferences ................................................................................................... 38
9 Z. [9 t5 f# J `/ ^; T3 Morphology of Human Embryonic Stem Cells and Induced
! f" k$ I8 S* ]7 Y t# Z# PPluripotent Stem Cells Cultured in Feeder
6 `6 _2 }+ j! A6 ?# \Layer-Free Conditions .............................................................................. 41
3 P7 b. h" A, ~8 Y3.1 Introduction ........................................................................................ 417 T( [( `9 j+ r& q* j8 v# O, N$ W
3.2 Materials for Feeder Layer-Free Culture of hPSCs ........................... 43
( c% E( Q& h S3.2.1 Matrix Preparation ................................................................. 43* T$ E/ Z$ K! a6 g* l- Z" z7 U
3.2.2 Culture Medium ..................................................................... 447 E" B6 j1 @) [( Z5 j! D* \5 y1 Y
3.3 Methods for hPSC Feeder Layer-Free Culture .................................. 44
! N2 W8 R; R- b0 K8 M' @3.3.1 Preparation of Matrix-Covered Plates ................................... 44+ c" ^+ [2 e9 {4 I- }
3.3.2 Splitting, Freezing, and Thawing hPSCs ............................... 455 H z4 J' D$ g# f, x V8 Q+ Q2 x
3.3.3 Adaptation of PSCs to Feeder-Free Culture .......................... 45" J4 j6 v+ B9 s6 v
3.3.4 Routine Culture of hPSCs ...................................................... 46+ h7 Q/ h) ` Z/ i E1 B
References ................................................................................................... 545 {+ F# E+ a$ b) x. z& z8 Q* ^
4 Morphology of Undifferentiated Human Embryonic' \7 S& s2 j' c4 O( r7 N
and Induced Stem Cells Grown in Suspension
2 a6 t% `" }9 J6 uand in Dynamic Cultures .......................................................................... 57
( D# k2 a/ K* V; Y- T7 G; p/ T& a# k4.1 Introduction ........................................................................................ 575 s0 d, s/ @( W3 b+ ?- K
4.2 Materials for Suspension Culture of hPSCs ...................................... 587 o9 ]6 y$ F1 \9 e1 F" Y
4.2.1 Culture Medium ..................................................................... 58# C8 y4 @7 Y* D/ k4 ]
4.2.2 Splitting Medium ................................................................... 59
' I: ?. G5 v, E$ O9 I( F4.2.3 Freezing Medium ................................................................... 59
" Q% N- }; C! Y4.3 Methods for Suspension Culture of hPSCs ....................................... 60
; j/ f0 t8 o8 r, J0 W/ a9 o# I- c* C4.3.1 Creating a hPSC Suspension Culture .................................... 607 { c" \+ f) v) Z1 t
4.3.2 Splitting hPSCs in Suspension ............................................... 604 ~& e! ?& U! \( I
4.3.3 Freezing hPSCs in Suspension .............................................. 62
+ t- }8 B' y, G# r! j5 l4.3.4 Thawing hPSCs in Suspension .............................................. 643 b% r: f2 G9 P) f3 @# c
4.3.5 Culturing hPSCs in a Dynamic System ................................. 64
9 T7 l3 i8 ~" `' U# I/ {4.3.6 Routine Culture of hPSCs in Suspension .............................. 65
' `: z3 s. c6 H" C' i* `References ................................................................................................... 71$ L( z) q3 f+ F$ y0 k5 B+ w' `
5 Differentiation of Pluripotent Stem Cells In Vitro:% M6 T" w5 V& v! ?8 @: M; g+ v* w
Embryoid Bodies ....................................................................................... 73
9 _* R; _8 l! q/ U3 Z5.1 Introduction ........................................................................................ 73
* d% F* p. w9 o5.2 Materials for EB Formation ............................................................... 75
U- q4 g4 d% f0 v' j9 _5.2.1 Culture Medium Supplemented with Serum ......................... 750 E; L8 v9 L# t
5.2.2 Splitting Medium Based on Collagenase ............................... 76
( n! ~# n' J( K4 _$ X p' `5.3 Methods for EB Formation and Culture ............................................ 76
P- R& I- [& _: [5.3.1 EB Formation ......................................................................... 76/ M5 D' C2 |; u/ z4 h# e9 x
5.3.2 Routine Culture of EBs .......................................................... 76, n( ^1 W" m; `4 V2 W2 J5 }# \
5.3.3 Culturing EBs in Spinner Flasks ............................................ 78
/ K3 S( t5 ]( e; M4 w6 MReferences ................................................................................................... 88, f! I+ c4 g0 L+ h: V) v+ a
6 Differentiation of Pluripotent Stem Cells In Vivo:
. O7 D0 v! G) h2 A( u: z3 |Teratoma Formation ................................................................................. 915 S; w3 k+ s/ O# e$ t0 ~5 K% k
6.1 Introduction ........................................................................................ 91% g1 k+ d& L2 g' R8 g5 M
6.2 Materials for Teratoma Formation ..................................................... 93
|9 D7 Z; p# D( F' J6.2.1 Culture Medium ..................................................................... 93
9 ` e, i! ] k7 y+ q( Y6.2.2 Syringe for Injecting Cells ..................................................... 93
1 \. o! b% Z! F3 n* ~' w. Q. Y5 f6.3 Formation of Teratomas ..................................................................... 934 Q' I% N1 L% n6 ]' X
6.3.1 Protocol for Teratoma Formation .......................................... 93
: c4 F" e* A2 i8 K, U+ o4 l+ i6.3.2 Routine Treatment of Mice and Teratoma ............................. 93
" o# h7 y6 ]/ j9 jReferences ................................................................................................... 103# \% R3 g* ]! `& F0 ~. U
7 Immunostaining ........................................................................................ 105/ [3 h/ }, F4 G* q% J5 o4 k8 R0 F
7.1 Introduction ........................................................................................ 105, S" M# n2 Q3 j" r8 A
7.2 Materials and Solutions for Immunostaining .................................... 1118 w( f) ^2 E! }/ U
7.2.1 Materials and Solutions for Immunohistochemistry' I a0 @ w* e; I; B/ K
of Paraffi n-Embedded Tissues ............................................... 111
8 r" O- g3 C& S- B7.2.2 Materials and Solutions for Immunofl uorescence ................. 111
6 b, V6 s' L+ @3 D6 I; Z v7.3 Immunostaining Procedures .............................................................. 112$ Y+ x- ]1 G8 B
7.3.1 Immunohistochemistry of Paraffi n-Embedded Tissues ......... 112+ }# f2 C v3 b( `
7.3.2 Immunofl uorescence of Cultured Cells ................................. 113, P/ T9 S5 z) n2 P9 `4 U
References ................................................................................................... 113
+ j# H7 w) T; X& I# [. a8 Karyotype and Fluorescent In Situ Hybridization
$ A( H- l, U; y, G$ ^Analysis of Human Embryonic Stem Cell and Induced
2 D% _& W8 G/ L2 T2 [Pluripotent Stem Cell Lines ..................................................................... 115/ H9 j) I% W1 Y$ O
8.1 Introduction ........................................................................................ 115& A' L6 I4 O+ H+ T, }( [3 ~! e
8.1.1 Karyotype Analysis ............................................................... 115
: \$ d' a7 t6 T4 y, _2 r n8.1.2 FISH Analysis ........................................................................ 121
; O5 l! _) u/ ?0 N d; y5 f8.2 Materials for Harvesting Cells for Karyotyping0 Q) g. r( n2 N1 a( d0 s2 v
and FISH Analysis ............................................................................. 124
/ \- M, P- f" |8.2.1 Reagents ................................................................................. 1240 @5 T' t. W6 l% O+ T2 T
8.2.2 Solutions ................................................................................ 1248 d2 b3 D4 x$ L
8.3 Procedure of Harvesting Cells for Karyotyping
* }" O A/ f: u/ D d; }and FISH Analysis ............................................................................. 1242 a. a, v+ y/ O9 C
References ................................................................................................... 126% O4 v( c o; I) C) S7 y6 Y, \6 M
9 Method for the Derivation of Induced Pluripotent
$ _) R/ e0 d8 aStem Cells from Human Hair Follicle Keratinocytes ............................ 127
/ T9 ?. w* N* i9.1 Introduction ........................................................................................ 127% g# U6 w! j& ? b) V
9.2 Materials ............................................................................................ 1296 O2 j& j6 E! m' L
9.2.1 NIH-3T3/293T Cells .............................................................. 129
) V+ W& i5 f) V( r! X9 Q8 G9.2.2 Keratinocyte Derivation from Plucked Hair Follicles ........... 129' V- m+ i0 W- L. q; K9 C
9.3 Methods ............................................................................................. 130! c) D1 q8 M, b+ l
9.3.1 NIH-3T3 and 293T Culture Methods .................................... 1305 d* g l3 F1 k; i
9.3.2 Keratinocyte Culture Methods ............................................... 132* O( U2 _) y- R
9.3.3 Preparation of the STEMCCA Virus for Infection ................ 133+ m8 z9 o; a- H$ R% C1 b, w
9.3.4 Derivation of iPSCs from Hair Keratinocytes ....................... 1341 v/ Q- f) Q& M0 Z
References ................................................................................................... 137 |
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发表于 2012-1-13 13:55
