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Protocols

ES Cells | iPS Cells | Vectors | ESC/iPSC Directed Differentiation Protocols

 

Embryonic Stem Cell Culture Protocols

  1. Mouse ESC Media with Serum
  2. Serum free ESC maintenance media
  3. ESC Passage
  4. Thawing and Freezing ES Cells
  5. Transduction of mESC with Lentivirus
  6. Making Mouse Embryonic Fibroblasts (MEFs)
  7. Shipping Tail Tips
  8. 2i Media (Feeder Free, Serum Free)

iPS Cell Protocols

  1. Human Dermal Fibroblast Isolation
  2. Shipping Live Culture Fibroblasts
  3. Isolation of Peripheral Blood Mononuclear Cells for Reprogramming
  4. Mouse iPS Cell Generation with STEMCCA
  5. Human iPS Cell Generation with hSTEMCCA
  6. Human iPS Cell Generation from Peripheral Blood with hSTEMCCA
  7. Southern Blot with human gDNA to screen for hSTEMCCA integration in hiPS cells
  8. Preparation of mouse tail tip fibroblasts

iPS Cell Core SOPs

  1. Preparing hESC Medium
  2. Preparing MEF Feeder Layer Plates
  3. Passaging hiPS Cells on Feeders using Collagenase
  4. Passaging hiPS Cells on Matrigel using Gentle Cell
  5. Passaging hiPS Cells on Matrigel using ReLeSR
  6. Isolation of Peripheral Blood Mononuclear Cells (PBMCs)
  7. Immunostaining of hiPS Cells for Pluripotency Markers
  8. Generation of iPS Cells from Human Peripheral Blood
  9. Freezing hiPS Cells Growing on MEF Feeders
  10. Freezing hiPS Cells Growing on Matrigel
  11. Thawing a Vial for Expansion and Banking (Stock and Distribution Vials)
  12. Thawing hiPS Cells onto Matrigel
  13. Thawing hiPSCs onto Feeders
  14. Excision of STEMCCA vector from hiPSCs by transient expression of Cre-recombinase
  15. Establishing iPSC lines from a reprogramming experiment

Vector Protocols

  1. Protocol 1: Lentivirus Packaging by 293T Transfection
  2. Protocol 2: Calculations for Transfection of 293T for Lenti/Retro Packaging
  3. Protocol 3: Titering Lentivirus on FG293 Cells
  4. Protocol 4: Titering Lentiviral Vectors by Southern Blot
  5. Protocol 5: Retrovirus Packaging
  6. Protocol 6: Primer Design for Cloning Genes into the pHAGE Lentiviral Backbone
  7. Protocol 7: Bacterial Storage and Glycerol Stocks
  8. Protocol 8: Lentiviral Transduction of Hematopoietic Stem Cells
  9. Protocol 9: Lentiviral Transduction of Alveolar Macrophages by Intratracheal Injection
  10. Protocol 10: Lentiviral infection of iAEC2s

**The Boston University Center for Regenerative Medicine endorses the International Stem Cell Registry (U.Mass), and The National Stem Cell bank (WiCell; University of Wisconsin). Detailed human ES Culture protocols and cell banks are maintained at each of these sites; click below to access their highly recommended protocol pages:

  1. International Stem Cell Registry
  2. National Stem Cell Bank; WiCell