Bacopa monnieri is used in traditional Ayurvedic medicine in India for centuries to control asthma, epilepsy and mental disorder. Various studies have shown that Bacopa is useful in controlling Alzheimer's disease, anxiety, and age-related cognitive decline. It is also used to improve mental function and memory, treat respiratory ailments, and heal stomach ulcers. The present study aimed to in vitro investigate the explants position and carbohydrate sources effect on the growth and development of Bacopa monnieri which were cultured in the MS nutrient medium supplemented with different concentrations (20, 30, 40 g\L) of the combination between the carbohydrates (sucrose, fructose, glucose) with the solid and the liquid media type and using stem of Bacopa plant containing of one node, two nodes and three nodes then cultured of two positions, horizontally and vertically method in the solid media. The incubation conditions were 25±2°C at 16 h/day photoperiod (2000-2500 lux). Glucose, 20 g\L, gave the best shoot number (148.75), shoot length (5.87), Growth value (4.25) and multiplication rate (15.00). Results showed that the best shoot number (17.00), Leaf number (76.00) and Node number (38.00) resulted from two nodes horizontally position. The best Shoot length (4.12) occurred due to one node horizontally position. The best root number (23.00) resulted from three node horizontally method.In vitro plantlets produced from rooting stage were transferred in plastic pots (8 cm diameter) containing peat, perlite and sand for acclimatization. The rooted plants were successfully acclimatized in different potting media and grew naturally in the greenhouse. The best planting medium for plant growth was peat moss +perlite mixture (2: 3).