Association of anxiolytic drugs diazepam and lorazepam, and the antiepileptic valproate, with heart defects—Effects on cardiomyocytes in micromass (MM) and embryonic stem cell culture

Evaluation of Bupropion Hydrochloride Developmental Cardiotoxic Effects in Chick Cardiomyocyte Micromass Culture and stem cell derived Cardiomyocyte Systems

“…The 3T3 fibroblast cell line is used to mimic maternal toxicity, but inhibition of fibroblast proliferation does not represent the in vivo situation and is omitted in number of studies (Anneelieke et al, 2007;Marx-Stoelting et al, 2009). In current study, 3T3 cell line is not used as done earlier by Ahir et al (2011). The mouse ES cell differentiation into cardiomyocyte using embryoid body (EB) formation assay resembles the in vivo cardiogenesis (Wei et al, 2005).…”

“…For ESDC system, the D3 mouse (Mus musculus) ES cells obtained from ATCC, Rockvile, USA (CRL-1934) were grown in undifferentiated state in the presence of leukemia inhibitory factor (Millipore-LIF1010). The ES cells were differentiated into cardiomyocytes (DMEM supplemented with 20% heatinactivated fetal bovine serum, 2 mM L-glutamine, 50 units/ml penicillin/50 g/ml streptomycin, 0.1 mM ␤-mercaptoethanol, and 1% X100 nonessential amino acids) using the hanging drop method in the presence of drug as described earlier by Ahir and Pratten (2011). In brief, after 3 days of hanging drop, the EBs were drained and further cultured for 2 days.…”

“…The cultured cardiomyocyte contractile activity in MM (24, 48, and 144 hr) and ESDC systems (days 10, 11, and 12) was recorded using light microscope. The numerical values were assigned to contractile activity scoring depending on the number of foci contracting compared to the control as described earlier (Memon and Pratten, 2009;Ahir et al, 2011). In brief, a score 3 is given if the entire well's foci are contracting like control, score 2 if numerous foci (not all) are contracting in a well, score 1 for few contracting foci (one or two), and zero for no contraction (Table 1).…”

“…In this study, BPN developmental cardiotoxic effects were evaluated using two in vitro systems; the chick cardiomyocytes micromass (MM) culture system and mouse embryonic stem cell derived cardiomyocyte (ESDC) system (Ahir et al, 2011;Shaikh Qureshi et al, 2014). The MM and ESDC system represents different stages of cardiac development.…”

“…The MM and ESDC system represents different stages of cardiac development. In MM system, the cardiomyocytes at the time of isolation were differentiated compared to ESDC system (Ahir et al, 2011;Shaikh Qureshi et al, 2014), where undifferentiated embryonic stem (ES) cells undergo spontaneous differentiation into contracting cardiomyocytes resembling the in vivo process of cardiac development (Wei et al, 2005). Evaluating the effects of BPN in these in vitro systems will enable us to understand its effects on cardiac development.…”