Λεπτομέρειες βιβλιογραφικής εγγραφής
| Τίτλος: |
Reprogramming Induced Pluripotent Stem Cell Lines from Frozen Buffy Coat Samples. |
| Συγγραφείς: |
Art J; Center for Molecular Medicine, University of Georgia; Interdisciplinary Neuroscience, University of Georgia; js.esquibel@yahoo.com., James C; Center for Molecular Medicine, University of Georgia; Department of Biochemistry and Molecular Biology, University of Georgia., Dalal B; Center for Molecular Medicine, University of Georgia; Department of Biochemistry and Molecular Biology, University of Georgia., Fantone K; Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia., Rada B; Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia., Felner EI; Division of Pediatric Endocrinology, Emory University School of Medicine., Gonzalez-Duarte A; Dysautonomia Center, New York University Langone Health., Michopoulos V; Department of Psychiatry and Behavioral Sciences, Emory University School of Medicine., Corneo B; Stem Cell Core, Columbia Stem Cell Initiative, Columbia University., Zeltner N; Center for Molecular Medicine, University of Georgia; Department of Biochemistry and Molecular Biology, University of Georgia; Department of Cellular Biology, University of Georgia. |
| Πηγή: |
Journal of visualized experiments : JoVE [J Vis Exp] 2026 Apr 10 (230). Date of Electronic Publication: 2026 Apr 10. |
| Τύπος έκδοσης: |
Journal Article; Video-Audio Media; Research Support, N.I.H., Extramural |
| Γλώσσα: |
English |
| Στοιχεία περιοδικού: |
Publisher: MYJoVE Corporation Country of Publication: United States NLM ID: 101313252 Publication Model: Electronic Cited Medium: Internet ISSN: 1940-087X (Electronic) Linking ISSN: 1940087X NLM ISO Abbreviation: J Vis Exp Subsets: MEDLINE |
| Imprint Name(s): |
Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]- |
| Ιατρικοί όροι (MeSH): |
Induced Pluripotent Stem Cells*/cytology , Blood Buffy Coat*/cytology , Cellular Reprogramming Techniques*/methods, Leukocytes, Mononuclear/cytology ; Humans ; Cryopreservation ; Cellular Reprogramming |
| Περίληψη: |
Human pluripotent stem cells (hPSCs) are a valuable tool for disease modeling. Further stem cells can be reprogrammed from adult somatic cells, called induced pluripotent stem cells (iPSC). iPSC technology allows for the evaluation of specific study participants and populations and ventures into personalized medicine. Blood is routinely taken and cryopreserved for research purposes. These samples are processed either as buffy coats, a blood sample containing white blood cells and platelets, or as peripheral blood mononuclear cells (PBMCs), which represent a more purified population of white blood cells without eosinophils, basophils, platelets, or red blood cells. Both are a readily available and relatively non-invasive source for reprogrammable somatic cells. Several reports detail reprogramming from PBMCs, whereas only one describes this process from frozen buffy coats. Recent experience revealed PBMC reprogramming protocols available in the literature and from manufacturers to be unsuccessful when applied to frozen buffy coat samples, necessitating the adaptations and troubleshooting strategies described here. For many researchers, who employ iPSC technologies, it is imperative to have thorough protocols with a high success rate, especially in cases, where patient samples may contain only few cells, are obtained in wide time intervals, are from limited participant pool, or are otherwise highly valuable. Here, checkpoints and troubleshooting strategies are identified to increase the chance of reprogramming human frozen buffy coats or purified PBMCs. Ultimately, this protocol will allow researchers to identify predictors of reprogramming success and strategize alternative approaches to improve the chances of successful iPSC derivation. |
| Grant Information: |
T32 GM142623 United States GM NIGMS NIH HHS; R01 MH115174 United States MH NIMH NIH HHS; R01 NS114567 United States NS NINDS NIH HHS |
| Entry Date(s): |
Date Created: 20260427 Date Completed: 20260716 Latest Revision: 20260727 |
| Update Code: |
20260728 |
| DOI: |
10.3791/70522 |
| PMID: |
42044032 |
| Βάση Δεδομένων: |
MEDLINE |