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Generation of Induced Pluripotent Stem Cells from Frozen Buffy Coats using Non-integrating Episomal Plasmids
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Developmental Biology
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JoVE Journal Developmental Biology
Generation of Induced Pluripotent Stem Cells from Frozen Buffy Coats using Non-integrating Episomal Plasmids

Generation of Induced Pluripotent Stem Cells from Frozen Buffy Coats using Non-integrating Episomal Plasmids

DOI:
10.3791/52885-v

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10:52 min

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June 05, 2015

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, , , , , , , , ,
1Center for Biomedicine,European Academy Bozen/Bolzano (EURAC), 2Laboratory of Medical Genetics,Fondazione IRCCS Ca´ Granda, Ospedale Maggiore Policlinico, 3Del E. Webb Center for Neuroscience, Aging & Stem Cell Research,Sanford-Burnham Medical Research Institute

Chapters

  • 00:05Title
  • 01:44Obtaining Peripheral Blood Mononuclear Cells (PBMNCs) from Buffy Coats
  • 02:46Culturing PBMNCs Obtained from Buffy Coats
  • 04:36Transfecting PBMNCs
  • 06:19Culturing Transfected PBMNCs on Mouse Embryonic Fibroblasts (MEFs)
  • 07:32Generating Induced Pluripotent Stem Cells (iPSC) Clones
  • 08:22Results: Cellular Characteristic from PBMNC to iPSC
  • 10:21Conclusion

Summary

Automatic Translation

Automatic Translation

Induced pluripotent stem cells (iPSCs) represent a source of patient-specific tissues for clinical applications and basic research. Here, we present a detailed protocol to reprogram human peripheral blood mononuclear cells (PBMNCs) obtained from frozen buffy coats into viral-free iPSCs using non-integrating episomal plasmids.

Tags

Induced Pluripotent Stem CellsIPSCsNon-integrating Episomal PlasmidsReprogrammingPeripheral Blood Mononuclear CellsPBMNCsFrozen Buffy CoatsRegenerative MedicinePatient-specificTransgene-free

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