Noninvasive Reporter Gene Imaging of Human Oct4 (Pluripotency) Dynamics During the Differentiation of Embryonic Stem Cells in Living Subjects

被引：9

作者：

Ahn, Byeong-Cheol ^{[1
,2
]}

Parashurama, Natesh ^{[1
,7
]}

Patel, Manish ^{[1
]}

Ziv, Keren ^{[1
]}

Bhaumik, Srabani ^{[3
]}

Yaghoubi, Shahriar Shah ^{[1
]}

Paulmurugan, Ramasamy ^{[1
,4
]}

Gambhir, Sanjiv Sam ^{[1
,4
,5
,6
]}

机构：

[1] Stanford Univ, Stanford Sch Med, James H Clark Ctr, MIPS,Dept Radiol,Div Nucl Med, Stanford, CA 94305 USA

[2] Kyungpook Natl Univ, Kyungpook Natl Univ Hosp, Sch Med, Dept Nucl Med, Taegu 700712, South Korea

[3] GE Global Res Ctr, Niskayuna, NY USA

[4] Canary Ctr Early Detect Canc, Palo Alto, CA USA

[5] Stanford Sch Med, Dept Bioengn, Stanford, CA USA

[6] Stanford Univ, Dept Mat Sci & Engn, Stanford, CA 94305 USA

[7] UCSF, Sch Med, Dept Obstet Gynecol & Reprod Sci, Inst Regenerat Med & Stem Cells, San Francisco, CA 94143 USA

来源：

MOLECULAR IMAGING AND BIOLOGY | 2014年 / 16卷 / 06期

关键词：

Embryonic stem cells; Pluripotent stem cells; Differentiation; Human Oct4; Reporter gene imaging; Gene networks; Molecular imaging; Oct4; Pluripotency; In vivo imaging; In vivo differentiation; SELF-RENEWAL; ANALYSIS REVEALS; EXPRESSION; NANOG; MICE;

D O I：

10.1007/s11307-014-0744-1

中图分类号：

R8 [特种医学]; R445 [影像诊断学];

学科分类号：

1002 ; 100207 ; 1009 ;

摘要：

Human pluripotency gene networks (PGNs), controlled in part by Oct4, are central to understanding pluripotent stem cells, but current fluorescent reporter genes (RGs) preclude noninvasive assessment of Oct4 dynamics in living subjects. To assess Oc4 activity noninvasively, we engineered a mouse embryonic stem cell line which encoded both a pOct4-hrluc (humanized renilla luciferase) reporter and a pUbi-hfluc2-gfp (humanized firefly luciferase 2 fused to green fluorescent protein) reporter. In cell culture, pOct4-hRLUC activity demonstrated a peak at 48 h (day 2) and significant downregulation by 72 h (day 3) (p=0.0001). Studies in living subjects demonstrated significant downregulation in pOct4-hRLUC activity between 12 and 144 h (p = 0.001) and between 12 and 168 h (p = 0.0003). pOct4-hRLUC signal dynamics after implantation was complex, characterized by transient upregulation after initial downregulation in all experiments (n = 10, p = 0.01). As expected, cell culture differentiation of the engineered mouse embryonic stem cell line demonstrated activation of mesendodermal, mesodermal, endodermal, and ectodermal master regulators of differentiation, indicating potency to form all three germ layers. We conclude that the Oct4-hrluc RG system enables noninvasive Oct4 imaging in cell culture and in living subjects.

引用

页码：865 / 876

页数：12