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Differentiation of 3T3-L1 pre-adipocytes 3T3-L1 pre-adipocytes cells were cultured with standard culture media (DMEM, 4.5 g/L glucose, L-glutamine, sodium pyruvate, 10% FBS, 1% antibiotic-antimycotic solution) and grown for 48 h before initiating differentiation using the manufacturer's suggested protocol and modified from previous published work.[18] Briefly, standard culture media was supplemented with scheduled addition of adipogenic agents [3-isobutyl-1methyl xanthine (IBMX), Sigma Aldrich

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I did once monthly, then once weekly injections here in clinics (after relying only on oral ssuoplementation )