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Chapters
- 00:06Assembling handing drops differentiation culture
- 00:08Collect undifferentiated ES cells
- 02:24Prepare materials
- 02:57Assemble hanging drop culture
- 04:34Collecting differentiated ES cells form hanging drops
- 06:05Starting EB culture
- 06:53EB dissociation and infection
- 11:25Preparation of ES cell-derived progenitors for FACS
- 15:18Transplantation
- 16:19Discussion
This protocol details the derivation of transplantable hematopoietic stem cells from mouse embryonic stem cells (ESC) and their subsequent injection into lethally irradiated recipient mice. Briefly, ESC are differentiated as embryoid bodies, which are then infected with retroviral HoxB4 and co-cultured with OP9 stromal cells and hematopoietic cytokines.
Tags
Stem CellsHematopoietic Stem CellsMurine Embryonic Stem CellsSelf-renewalDifferentiated ProgenyPluripotentDifferentiative AbilityClinically Useful Cell TypesVideo ProtocolHematopoietic MalignanciesHematopoietic DisordersHSC TherapyAlternative Source Of HSCBaselineHuman ESC IsolationEmbryoid Bodies (EBs)Serum Pre-screenedHematopoietic DifferentiationRetroviral HoxB4OP9 StromaBone Marrow Stromal Cell LineCytokines
