title: Efficient generation of integration-free ips cells from human adult peripheral blood using BCL-XL together with Yamanaka factors.
abstract: The ability to efficiently generate integration-free induced pluripotent stem cells (iPSCs) from the most readily available source-peripheral blood-has the potential to expedite the advances of iPSC-based therapies. We have successfully generated integration-free iPSCs from cord blood (CB) CD34(+) cells with improved oriP/EBNA1-based episomal vectors (EV) using a strong spleen focus forming virus (SFFV) long terminal repeat (LTR) promoter. Here we show that Yamanaka factors (OCT4, SOX2, MYC, and KLF4)-expressing EV can also reprogram adult peripheral blood mononuclear cells (PBMNCs) into pluripotency, yet at a very low efficiency. We found that inclusion of BCL-XL increases the reprogramming efficiency by approximately 10-fold. Furthermore, culture of CD3(-)/CD19(-) cells or T/B cell-depleted MNCs for 4-6 days led to the generation of 20-30 iPSC colonies from 1 ml PB, an efficiency that is substantially higher than previously reported. PB iPSCs express pluripotency markers, form teratomas, and can be induced to differentiate in vitro into mesenchymal stem cells, cardiomyocytes, and hepatocytes. Used together, our optimized factor combination and reprogramming strategy lead to efficient generation of integration-free iPSCs from adult PB. This discovery has potential applications in iPSC banking, disease modeling and regenerative medicine.
authors.affiliation: Department of Medicine, Loma Linda University, Loma Linda, California, United States of America.
authors.forename: Rui-Jun
authors.initials: RJ
authors.lastname: Su
authors.forename: David J
authors.initials: DJ
authors.lastname: Baylink
authors.forename: Amanda
authors.initials: A
authors.lastname: Neises
authors.forename: Jason B
authors.initials: JB
authors.lastname: Kiroyan
authors.forename: Xianmei
authors.initials: X
authors.lastname: Meng
authors.forename: Kimberly J
authors.initials: KJ
authors.lastname: Payne
authors.forename: Benjamin
authors.initials: B
authors.lastname: Tschudy-Seney
authors.forename: Yuyou
authors.initials: Y
authors.lastname: Duan
authors.forename: Nancy
authors.initials: N
authors.lastname: Appleby
authors.forename: Mary
authors.initials: M
authors.lastname: Kearns-Jonker
authors.forename: Daila S
authors.initials: DS
authors.lastname: Gridley
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: K-H William
authors.initials: KH
authors.lastname: Lau
authors.forename: Xiao-Bing
authors.initials: XB
authors.lastname: Zhang
mesh_terms: D000328:Adult; D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001773:Blood Cells / Q000166:cytology* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D019070:Cell Lineage; D065150:Cellular Reprogramming; D005312:Fetal Blood / Q000166:cytology; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D051020:bcl-X Protein / Q000378:metabolism*
chemical_list: D018952:Antigens, CD34; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors; D051020:bcl-X Protein