Background & Aim Induced pluripotent stem cells (iPSCs), which can be generated from patients’ somatic cells and grown in cultures maintaining their pluripotency and possessing the unlimited self-renewal capability, have been utilized for developing cell therapeutic strategies, dissecting disease mechanisms and validating drug response. Recent studies imply pluripotent stem cells (PSCs) possess a unique metabolic features and require specific metabolites in maintaining their cell fate. In the current work, we have deciphered the specific metabolites required for maintaining self-renewal expansion of iPSCs. Methods, Results & Conclusion Utilizing RNA-Seq analysis, we showed that, in comparing with fibroblast, ESCs and iPSCs were found to retain the expression of critical enzymes involved in Krebs cycle. We then determined if the specific bioenergetics adaptation in cultures can affect pluripotency acquirement or maintenance. Oct4-GFP(+) iPSCs derived mouse fibroblast were seeded in medium containing either 4500mM Glucose (high glucose) with or without Oligomycin (mitochondrial ATPase inhibitor), 500mM Glucose (low glucose) and 0mM Glucose (no glucose) with or without pyruvate (to be utilized in TCA cycles). We found that depletion of glucose resulted in reduced growth of Oct4-GFP(+) iPSC colonies. In contrast, addition of pyruvate could rescue the growth of Oct4-GFP(+) iPSC colonies suggesting the retaining mitochondrial respiration may aid self-renewal expansion of pluripotent stem cells. The further analysis revealed that mouse and human ESCs/iPSCs had evaluated level of GLUT5 & KHK suggesting pluripotent stem cells can utilize fructose efficiently. We further validated that fructose substitution was sufficient to maintain the self-renewal growth of ESCs/iPSCs. Importantly, fructose substitution was found to enhance generation of iPSCs from Yamanaka factor mediated fibroblast reprogramming possibly via selectively suppressing the growth of partial reprogrammed cells and upregulating alpha 2,6-sialylation as sialyltransferase ST6Gal1 is known to play an crucial role in regulating pluripotency. In summary, the current work elucidated that the specific bioenergetics adaptation in cultures linking to pluripotency maintenance and identified ESCs/iPSCs can utilize fructose efficiently. Fructose substitution can therefore be used to enhance derivation of iPSCs.