Basic Cell Culture Protocols
THIRD EDITION
Edited by
Cheryl D. Helgason
Cindy L. Miller
Contents
1.Culture of primary Adherent Cells and a Continuously Growing Nonadherent Cell Line 1
2.Detection of Mycoplasma Contaminations 13
3.Eradication of Mycoplasma Contaminations 25
4.Authentication of Scientific Human Cell Lines: Easy-to Use DNA Fingerprinting 35
5.Cytogenetic Analysis of Cell Lines 51
6.Human and Mouse Hematopoietic Colony-Forming Cell Assays 71
7.Isolation and Culture of Murine Macrophages 91
8.Isolation and Culture of Human Macrophages 105
9.Development of T-Lymphocytes in Mouse Fetal Thymus Organ Culture 117
10.In Vitro Generation of Lymphocytes From Embryonic Stem Cells 135
11.Hematopoietic Development of Human Embryonic Stem Cells in Culture 149
12.Generation of Murine Stromal Cell Lines: Models for the Microenvironment of the Embryonic Mouse Aorta-Gonads-Mesonephros Region 163
13.Culture of Human and Mouse Mesenchymal Cells 173
14.Isolation, Purification, and Cultivation of Murine and human Keratinocytes 187
15.Isolation and Culture of Primary Human Hepatocytes 207
16.Primary Kidney Proximal Tubule Cells 231
17.Enzymatic Dissociation and Culture of Normal human Mammary Tissue to Detect Progenitor Activity 249
18.Generation and Differentiation of Neurospheres From Murine Embryonic Day 14 Central Nervous System Tissue 265
19.Isolation and Culture of Skeletal Muscle Myfibers as a Means to Analyze Satellite Cells 281
20.Adult Ventricular Cardiomyocytes: Isolation and Culture 305
21.Isolation and Culture of Primary Endothelial Cells 315
22.Studying Leukocyte Rolling and Adhesion in Vitro Under Flow Conditions 331
23.Isolation and Characterization of Side Population Cells 343
24.Scalable Production of Embryonic Stem Cell-Derived Cells 353