Index

5-Bromo-2'-deoxyuridine, see

Immunofluorescence, DNA replication

5-Methylcytosine, 129, 251, see also DNA methylation and DNA demethylation antibody staining of, 134-135

6-Thioguanine, see Selection, HPRT

Activation of reconstructed oocytes, see Embryo, activation of NT mouse Actinomycin D, 230 Adipose tissue, 35-38 Aging, see Senescence Alkaline phosphatase, 182, 199, 210 histochemistry of, 218-219, 222 Anesthesia, 9, 20 Animal cap, 4

Animal pole, see Animal-vegetal axis Animal-vegetal axis, l Ascites, 210, 221 Avidin, see Immunohistochemical analysis

ß-geo neo/LacZ transgene, see Genetic markers, ß-geo Biotin, see Immunohistochemical analysis.

Bisulfite mutagenesis, 251-252, 255-256, see also DNA methylation Blastocyst, 131, 181, 182, 209

BMP, see Bone morphogenetic protein

Bone marrow, 35-36, 151 Bone morphogenetic protein, 182 BrDU, see Immunofluorescence, DNA replication

Cell counting, see Hemocytometer Cell cycle, 231 arrest, 150 G0 phase, 21, 93 G1 phase, 182 G1/S transition, 175 M phase, 226 Celltracker dyes, see Vital stains Centromere, 240

Chromatin, 139, 261-262, 273-274, 285-286 immunoprecipitation assay, 261268, 273-274, 278-280, 318321

ChlP assay, see Chromatin, immunoprecipitation assay Chromosomes, 5, 7, 12, 60, 131, 150, see also Telomeres abnormal, 174, 235 spreads of, 169-170 Cloning (animal), 25, 59, 131, 152, 225 Clones, defects in, 149-150, 152, 174,

230-231, 251 COCs, see Cumulus oocyte complexes Colcemid, 62, 65, 169, 175, see also

Microcells CpG dinucleotides, 129, 240, 251 Crosslinking, 146, 263-264, 276, 280281, 316, 318-319, 323, see also Fixation

Cumulus cells, see also Nuclear donor, cumulus cells as removal from sheep oocytes, 17-18 Cumulus oocyte complexes sheep, 11, 16, 21 mouse, 21, 28 Cytochalasin B, 62, 117, 124, see also Nuclear transfer, enucleation

DAPI, 124, 175, 216 Dense fibrillar component (DFC), 226228, see also Immunofluorescence, nucleolar; Immunogold; DNA immunocytochemistry; Nucleolus Developmental Studies Hybridoma

Bank, 197, 209 DFC, see Dense fibrillar component Differentiation, 101, 109, 151, 181183, 189-193, see also Embryoid body adipogenesis, 44 antibody markers of, 197 cardiogenesis, 189, 199 endodermal, 191 hepatic, 191, 199 matrices for, 199 neurogenesis, 189-191, 199 osteogenesis, 44 pancreatic, 191, 199 RNA expression markers of, 192-193 DMS footprinting, 288-289 DNA demethylation, 130-131, 251252, see also DNA methylation; 5-methylcytosine DNA immunocytochemistry, 234-235 DNA methylation, 129-131, 239-240, 251-252, see also 5-Methylcytosine; DNA demethylation-specific PCR, 243-245, 252-258

DNAse I footprinting, 289-290 DNA synthesis, 1, 76, 139, 144, 274, see also Immunofluorescence, DNA replication Dnmt1, 129 Dolly, 12, 25, 152, 174

EC cells, see Embryonic carcinoma cells

Ectoderm, 181-183, 199 embryonic, 209 extra-embryonic, 208 EG cells, Embryonic Germ cells Eggs, see Oocytes Electrofusion in nuclear transfer, 20 Electrophoresis, see Pulsed-field gel electrophoresis; Polyacrylamide gel electrophoresis ELISA, 198 Embryoid body, 182 preparation of, 189, 199 Embryonic carcinoma cells, 208, 210, 213

Embryonic germ cells, 68, 181, 208210, 213

Embryonic stem cells, 74-75, 181-183, 207-209, see also Differentiation; Oct-4 characteristics of, 182 culture of, 85-87, 188-189, 198199

feeder cells for, 74, 188, 222 markers of, 209-211 as nuclear donors, 31 nuclear reprogramming activity of, 68, 82 passaging, 75 Embryos activation of NT mouse, 30-31, 32, 124

antigens of, see Stage-specific embryonic antigens bovine, 228, 230-231, 240 cleavage failure in NT, 31 culture of NT reconstructed mouse, 31, 124-125 sheep, 20

DNA methylation status of, 239-240 isolation of DNA from, 241, 254255

isolation of preimplantation, 133 rabbit, 228, 230 transfer of, 20-21, 31 Endoderm, 181-183, 208

visceral, 209 Endothelial cells, 41 End replication problem, 150 Enzyme-linked immunosorbent assay, see ELISA Epiblast, see Inner cell mass Epigenetics, v, 67-68, see also Nuclear reprogramming regulatory marks, 129, 139, 285, 315 ES cells, see Embryonic stem cells Euchromatin, 131 Ewes, see Sheep, preparation for embryo transfer Extinction, vi

Extracellular matrix, 44, 95, 199

FACS, see Flow cytometry FC, see Fibrillar Center Fertilization, 67, 130, 133, 235 Fetal calf serum, 74, 199 Fibrillar center, 226, 228, see also

Nucleolus Fibroblasts, see Embryonic stem cells, feeder cells for FISH, Y chromosome, 64, see also Immunofluorescence, telomeres

Fixation, see also Crosslinking. of embryos 134 of cells, 143, 144, 146 Flow cytometry 84-85, 89-92, 214,

216-217 Fluorescence, 125 filters, 135

fluorochromes, 135, 144-145, 320 Formaldehyde, 316, see also

Crosslinking; Fixation Fusion, 47-48, 53, 59-60, 67-69, 81-85 electrofusion of cells, 75-76, 88-89 hybrids, 48, 82

culture of, 77, 89 monitoring of, 92 in nuclear transfer, see Nuclear transfer polyethylene glycol-induced, 51-52,

63-64 virus-induced, 69 in vivo, 47, 68

G418, see Selection, G418, see also

Mouse, strains, ROSA26 Gastrulation, 6 GC, see Granular component Genetic markers, 7, 115-116

P-geo, 76, see also Selection, G418;

Mouse, strains, ROSA26 GFP, 7, 94, 115 Geneticin, see Selection, G418; Mouse, strains, ROSA26 Genomic imprinting, 68, 129 Germinal vesicle, 6-7 GFP, see Genetic markers, GFP Globins, 48, 267 Glucose, 200

Granular component, 226, 228-230, see also Nucleolus Green fluorescent protein, see Genetic markers, GFP

HAT medium, see Selection, HPRT, HAT

HDACs, see Histone, deacetylases Hemocytometer, 50 Hemoglobin switching, 48 Heterochromatin, 131, 174 Heterokaryon, 47, 81 Histone, 139, 261, 273, see also Chromatin code, 139, 274, 315 deacetylases, 273 N-terminal tail modifications of, 68, 139-140, 262, 273, 321, 323 immunostaining for, 143-144 marks of active loci, 321 Homokaryon, 85 Hprt gene, see Selection, HPRT Hybrids, see Fusion, hybrids Hygromycin B, see Selection, hygromycin B

Immunofluorescence, 140 cells, 213-218 chromatin, 140, 143-144 controls for, 147, 213 differentiation, 195-197, 200 DNA methylation, 131, 134-135 DNA replication, 144 nucleolar, 232-233 SSEAs, 213-218 telomeres, 169-171, 175-176 Immunoglobulin, see Monoclonal antibody Immunogold, 233-234 Immunohistochemical analysis, 219-221 Immunostaining, see DNA

Immunocytochemistry;

Immunofluorescence;

Immunogold;

Immunohistochemical analysis Imprinting, see DNA imprinting

Inner cell mass, 12, 181, 207, 209 Insulin, 200 Interleukin-6, 182

LacZ, see Genetic markers, ß-geo Leukemia inhibitory factor, 182, 198, see also Embryonic stem cells, culture of

LIF, see Leukemia inhibitory factor, see also Embryonic stem cells, culture of Lipoaspiration, 37 Liposome, 52 Lysolecithin, 8

Magnetic cell sorting, 40-41 Meiosis, 1

meiotic spindle, 5 MEK/ERK signalling pathway, 182 Mesenchyme, 35 Mesoderm, 35, 44, 181-183 Metaphase, 5, 12, 25, 115, 122, 169, 175, 226, see also Chromosomes, spreads of Microcells, 60, 65

preparation of, 62-63 Micrococcal nuclease, see MNase Microinjection failure, 31-32 Microinjector, 14, 26 Micromanipulator, 14, 26 Microscope, 14-15, 26, 133 Microscopy confocal, 200 electron, 232

fluorescence, 41-42, 133, 176, see also Immunofluorescence phase contrast, 52-53 Mitotic clock, 150 MNase, 317, 324

footprinting, 289 Monoclonal antibody, 209-210, 221

Mouse, 25, 125, see also

Superovulation of mice culture of cloned embryos, 31 strains, 28, 85 CBA, 85 D2, 125 OctNeo, 85, 93 ROSA26, 72, 76 Murine embryonic fibroblasts, see

Embryonic stem cells, feeder cells for

Nanog, 82, 182

neo/LacZ transgene, see Genetic markers, P-geo; see also Selection; G418; Mouse, strains, ROSA26 Neomycin, see Selection, G418 Neuroectoderm, see Ectoderm Nuclear donor, v amphibian, 3-4, 6 cumulus cells as, 29, 116 ES cells as, 31

preparation of, 19-20, 29, 120 Nuclear reprogramming, v-vi, 47-48, 81-82, 99-101, 130-131 cellular extract-induced, 100-109 in cloning, 68, 150, 173-174, 251 fusion-induced, see Fusion nuclear transfer-induced, see Nuclear transfer

Nuclear transfer, v-vi, 1-9, 11-22, 2532, see also Cloning (animal); Clones, defects in to amphibian oocytes, 6-7 to enucleated amphibian eggs, 3-5 to mouse oocytes, 30, 120-124 pipets for, 3, 21, 30, 119, 127 to sheep oocytes, 20 Nucleolar organizing region (NOR), 226, see also Nucleolus

Nucleolar precursor body (NPB), 227230, see also Nucleolus Nucleolus, 225-231

markers of, 227 Nucleosomes, 273, 285-286, 315

footprinting of in vivo, 288-307, 309 Nucleus, 152, 226, see also Nuclear transfer counting, 323

isolation of, 109-110, 112, 296297

derivative transgenes, 93, 115-116 Oocytes, v-vi, 1, 11-12, 17, 67-68, 116, 158, 227 amphibian, enucleation of, 5-6 DNA from, 240-245

injection of, see Nuclear transfer, to enucleated amphibian eggs and to amphibian oocytes lysis of, see Microinjection failure mouse, enucleation of, 29-30, 124 injection of, see Nuclear transfer, to mouse oocytes isolation of, 28-29, 120 sheep, enucleation of, 18-19

injection of, see Nuclear transfer, to sheep oocytes isolation of, 15-16 surface markers, 209

PCR, see also DNA methylation, demethylation-specific PCR; Chromatin, immunoprecipitation assay of GC-rich sequences, 281 primer design, 253, 258, 307 primer optimization, 307-308 real-time, 162-164, 172-173, 320321

Permeabilization of cells, 143, 147, see also Streptolysin O PGCs, see Primordial germ cell Piezo-electric injection, 25, 30, 32, 116,

122-124, 127 Polar body, 11, 18-19 Polyacrylamide gel electrophoresis, 161, 268, 270, 306-307, 313 Polyethylene glycol, 53, 54, 59, 64, see also Fusion, polyethylene glycol-induced Polykaryon, 67

Primary embryonic fibroblasts, see

Embryonic stem cells, feeder cells for

Primordial germ cell, 68, 181, 207-210, 218

Pronucleus, 7, 9, 31, 67 Pulsed-field gel electrophoresis, 175

Reporter gene, see Genetic markers Ribosomes RNA, 226, 231

synthesis, inhibition of, 230 SCNT (Somatic cell nuclear transfer), see Nuclear transfer; Cloning (animal); Clones, defects in Selection, 65, 94

G418, 77, 87, see also Genetic markers, $-geo; Mouse, strains, ROSA26 HPRT, 60-62, 76, 93, 95 6-thioguanine, 65 HAT, 77, 87 HT, 87, 95 hygromycin B, 61-62 Senescence, 44, 151-152, 174 Serum, bovine, see Fetal calf serum Sheep, 11-12

preparation for embryo transfer, 16 repetitive DNA sequences in, 246-247

SINEs, see Sheep, repetitive DNA

sequences in Sodium bisulfite, see Bisulfite mutagenesis Sp1 transcription factor, 274 Sperm, 130, 174 Spermatogenesis, 151, 174 Spindle-chromosome complex, see

Oocytes, enucleation of mouse SSEA, see Stage-specific embryonic antigens

Stage-specific embryonic antigens, 182, 207, 209-211, see also Immunofluorescence, SSEAs Stem cells, see also Embryonic stem cell stromal stem cell, 35-44 culture of, 42-44 surface markers of, 36 Streptolysin O, 8, 105-106, 112 Superovulation of mice, 28, 120, 125, 133, 136

Telomerase, 149-151, 274

quantification of activity, 157-164 Telomeres, 149-152, 165-176, see also Immunofluorescence, telomeres in clones, 152 measurement of, 165-173 Teratocarcinoma, 182, 208 Teratoma, 68, 182, 222 Tetraploidy, 1 Thymocytes, 93

isolation of, 74-75, 88 Toxicity assay, 104-105 Transdifferentiation, vi, 47, 68, 115 TRAP assay, see Telomerase, quantification of activity Trypan blue, 8-9 Tukey simultaneous test, 246

Vegetal pole, see animal-vegetal axis Vital stains, 93-94 Vitelline membrane, 8

X Chromosome reactivation, 68 Xenopus, 1-9 X-gal staining, 125

Zona pellucida, 18, 29, 122, 158, 170

Diabetes 2

Diabetes 2

Diabetes is a disease that affects the way your body uses food. Normally, your body converts sugars, starches and other foods into a form of sugar called glucose. Your body uses glucose for fuel. The cells receive the glucose through the bloodstream. They then use insulin a hormone made by the pancreas to absorb the glucose, convert it into energy, and either use it or store it for later use. Learn more...

Get My Free Ebook


Post a comment