High–efficiency generation of induced pluripotent stem cells from human foreskin fibroblast cells using the Sagunja-tang herbal formula

Abstract

BackgroundSagunja-Tang (SGT-4) is a traditional herbal formula in Korean medicine that is used to treat anti-metabolic syndrome, and has antioxidant activity. In this study, we evaluated the effects of SGT-4 on the formation efficiency of induced pluripotent stem cells (iPSCs) from human foreskin fibroblasts (HFFs) by four reprogramming transcription factors: Oct4, Sox2, KIf4, and c-Myc (OSKM).MethodsSGT-4 contained four different herbal medicines that are composed of Ginseng Radix, Glycyrrhizae Radix et Rhizoma, Atractylodis Rhizoma Alba, and Poria Sclerotium. The composition of SGT-4 was analyzed by high-performance liquid chromatography (HPLC). HFFs were transfected with episomal vectors contained by four OSKM. Western blotting, RT-PCR, immunofluroescence, and in vitro differentiation were used to assess the pluripotency of the iPSC cells.ResultsSGT-4 exhibited antioxidant activity against the generation of intracellular reactive oxygen species (ROS) as well as promoted the activation of superoxide dismutase 1 (SOD1), catalase, gluthathione peroxidase 1 (GPX1), and glutathione (GSH). Moreover, the ATP level was not significantly fluctuated depending on the concentration of SGT-4 in the hiPSCs.ConclusionOur results indicate that the SGT-4, herbal formula significantly increases the efficiency of human iPSC generation via the transcription factors (Oct4, Sox2, KIf4, and c-Myc).

Prunellae Spica Extract Suppresses Teratoma Formation of Pluripotent Stem Cells through p53-Mediated Apoptosis

“…As these residual undifferentiated cells in CPTs are tumorigenic and can form teratomas at ectopic sites after in vivo transplantation, complete differentiation or selective elimination of residual undifferentiated cells in CTPs prior to transplantation is critical for clinical application [5,6]. Several methods for selective removal of undifferentiated iPSCs from a population of differentiated CTPs have been reported, such as modification of cell culture conditions, use of hiPSCs-specific sorting antibodies and chemical inhibitors, and introduction of suicide genes into hiPSCs [7][8][9][10]. However, none of these approaches are clinically applicable for regenerative therapy because of concerns regarding specificity, safety, and cost.…”

“…In addition, the AMP-activated protein kinase (AMPK) agonist, metformin, which specifically disrupts the cancer stem cell compartment in multiple cancers, limited the tumorigenicity of teratoma-initiating iPSCs without interfering with their pluripotency [35]. Small molecules such as PluriSin#1 (an inhibitor of stearoyl-CoA desaturase), quercetin and YM155 (inhibitors of survivin), dinaciclib (a CDK inhibitor), and cardiac glycosides have been shown to induce selective cell death of undifferentiated iPSCs and efficiently prevent teratoma formation [9,36,37]. However, all of these molecules have been investigated only in preclinical studies, and have not yet been approved for clinical use.…”

“…In many studies, herbal medicines have been shown to be useful adjuvants for the management of intractable diseases because of their high efficacy, minimal toxicity, and multi-modal activities [8]. Our research group recently reported that Sagunja-tang, a traditional Korean herbal formula, significantly increased the efficiency of hiPSC generation by reprogramming human foreskin fibroblasts using transcription factors (Oct4, Sox2, Klf4, and c-myc) [9]. However, herbal medicines that reduce the teratoma-forming activity of undifferentiated hiPSCs have not yet been reported.…”

Episomal Induced Pluripotent Stem Cells: Functional and Potential Therapeutic Applications

“…The human cells reported to have been successfully reprogrammed into EiPSCs include fibroblasts, epithelial cells, keratinocytes, mononuclear cells from adult peripheral blood, cord blood cells, amniotic fluid stem cells, mesenchymal stromal cells, lymphoblasts, lamina propria progenitor cells from oral mucosa, and urothelial cells obtained from urine. A summary of these reported human EiPSC sources is shown in Table 1 8,20,21,32 –43,47 –52,55,56,58,60 …”

Effects of the Extracts from Fruit and Stem of Camellia japonica on Induced Pluripotency and Wound Healing

“…Considerable efforts have been made to improve the efficiency and safety of somatic cellular reprogramming to generate genetically unmodified hiPSCs for clinical applications. Various small molecules, including epigenetic modifiers, metabolic regulators, and antioxidants, and herbal decoction Sagunja-Tang, have been identified that are involved in the reprogramming process and promote the efficiency of iPSC generation [ 13 , 14 , 15 , 16 ]. In particular, chemically induced iPSCs (CiPSCs) can be generated from mouse somatic cells using a combination of seven small molecules, including VC6T (VPA, CHIR99021 a GSK inhibitor, 616452 and Tranylcypromine a monoamine oxidase inhibitor) plus forskolin (a cAMP agonist) and 3-deazaneplanocin A (DZNep, a global histone methylation inhibitor) but not from human cells [ 17 ].…”