Basic Cell Culture Protocols

ISBN-13: 9781627031271 / Angielski / Twarda / 2012 / 550 str.

Cheryl D. Helgason; Cindy L. Miller

Opis

At some point in their careers, virtually every scientist and technician, as well as many medical professionals, regardless of their area of specialization have a need to utilize cell culture systems. Updating and significantly expanding upon the previous editions, Basic Cell Culture Protocols, Fourth Edition provides the novice cell culturist with sufficient information to perform the basic techniques, to ensure the health and identity of their cell lines, and to be able to isolate and culture specialized primary cell types. The intent of this extensive volume is to generate a valuable resource containing clear methodologies pertinent to current areas of investigation, rather than attempting to educate cell culturists on specific cell types or organ systems. Written in the highly successful Methods in Molecular Biology , chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Comprehensive and up-to-date, Basic Cell Culture Protocols, Fourth Edition compiles the essential techniques needed to approach this vital laboratory activity with full success.

Spis treści

1. Detection of Mycoplasma Contaminations

2. Eradication of Mycoplasma Contaminations

3. STR DNA Typing of Human Cell Lines: Detection of Intra- and Interspecies Cross-Contamination

4. Classical and Molecular Cytogenetic Analysis

5. Fluorescent In Situ Hybridization (FISH) of DNA Probes in the Interphase and Metaphase Stages of the Cell Cycle

6. The Development of T Lymphocytes in Fetal Thymus Organ Culture

7. Generation, Isolation, and Engraftment of In Vitro-Derived Human T Cell Progenitors

8. In Vitro Generation of Human T Regulatory Cells: Generation, Culture, and Analysis of FOXP3-Transduced T Cells

9. Simultaneous Cloning and Selection of Hybridomas and Transfected Cell Lines in Semisolid Media

10. Isolation and Characterization of Mouse Side Population Cells

11. Stem Cell Identification by DyeCycle Violet Side Population Analysis

12. Isolation and Characterization of Cancer Stem Cells In Vitro

13. Ex Vivo Differentiation of Cord Blood Stem Cells into Megakaryocytes and Platelets

14. Generation and Characterization of Murine Alternatively Activated Macrophages

15. Human Long Term Culture Initiating Cell (LTC-IC) Assay

16. Long Term Culture Initiating Cell (LTC-IC) Assay for Mouse Cells

17. Colony Forming Cell Assays for Human Hematopoietic Progenitor Cells

18. Studying Leukocyte Recruitment Under Flow Conditions

19. Generation and Establishment of Murine Adherent Cell Lines

20. Isolation, Enumeration, and Expansion of Human Mesenchymal Stem Cells in Culture

21. Isolation and Culture of Mesenchymal Stem Cells from Mouse Compact Bone  

22. Generation of a Pool of Human Platelet Lysate (pHPL) and Efficient Use in Cell Culture

23. In Vitro Methods to Culture Primary Human Breast Epithelial Cells

24. Human Prostate Epithelial Cell Cultures

25. Enzymatic Dissociation, Flow Cytometric Analysis, and Culture of Normal Mouse Mammary Tissue

26. Isolation and Characterization of Human Hair Follicle Epithelial Cells

27. Co-Cultivation of Human Oral Keratinocytes and Human Osteoblast-Like Cells

28. Isolation and Culture of Skeletal Muscle Myofibers as a Means to Analyze Satellite Cells

29. Hepatic Differentiation of Embryonic Stem Cells by Murine Fetal Liver Mesenchymal Cells

30. Methods to Culture, Differentiate, and Characterize Neural Stem Cells from the Adult and Embryonic Mouse Central Nervous System

31. Feeder-Independent Culture Systems for Human Pluripotent Stem Cells

32. Formation of Embryoid Bodies from Human Pluripotent Stem Cells Using AggreWell™ Plates

33. Techniques in Embryoid Body Formation from Human Pluripotent Stem Cells