Of biologically active B-complex vitamins, vitamins B6, B9 and B12 has been reported to influence maintenance of red blood cells, stem cells, nervous and immune systems, tissue regeneration and wound healing. However, the collective effects of restrictions/supplementations of vitamins B6, B9 and B12 on growth, vasculogenesis and senescence of BG01V human embryonic stem cell (hESC) derived human embryoid bodies (hEBs) is not understood. This study was proposed to understand the collective effects of restrictions/supplementations of vitamins B6, B9 and B12 on growth, vasculogenesis and senescence of hEBs. The collective effect of four different media conditions like deficient (0%), marginal (30%), adequate (70%) and supraphysiological levels of vitamins B6, B9 and B12 restrictions/supplementations on growth and viability of hEB derived vasculogenesis were studied by photometric analysis. Morphology and senescence of hEB derivatives were examined and compared using inverted microscopy and scanning electron microscopy. Out of four different media conditions like 0%, 30%, 70% and supraphysiological levels of vitamins B6, B9 and B12, media supplied with adequate level of vitamins B6, B9 and B12 to hESCs grown on 100% restriction were able to resume all activities at normal. The BG01V hESC-hEBs grown without vitamins B6, B9 and B12 showed cell disintegration, cell floating, SA-β-gal expression, cell lose, decrease size, number and metabolic activity. The hEB derived vascular cell like structures grown on similar conditions showed compromised self-renewal capacity, vulnerable growth and early senescence. The initial damages due to restrictions may be reversed by adequate (70%) supply of vitamins B6, B9 and B12; however reversal of damage may not be 100%.