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. Author manuscript; available in PMC: 2022 Jan 1.
Published in final edited form as: Methods Mol Biol. 2021;2158:171–186. doi: 10.1007/978-1-0716-0668-1_13

Table 1 –

Cardiogenic cell types, sources, and culture media used for maintenance of engineered cardiac tissues generated by the frame-hydrogel method.

Cell Type Cell Source Culture Media
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) Differentiated from monolayers of human iPSCs using small molecule modulation of the Wnt pathway [24]. Purified using metabolic selection [25]. Days 0–7 days of culture:
RPMI-1640, 2% B-27 supplement, 2 mg/mL aminocaproic acid, 50 ug/mL ascorbic acid 2-phosphate, 1% penicillin-streptomycin, 1% non-essential amino acids, 1% sodium pyruvate, 0.45μM 1-thioglycerol
Days 7–21 of culture:
Low-glucose DMEM, 5% fetal bovine serum, 2 mg/mL aminocaproic acid, 50 ug/mL ascorbic acid 2-phosphate, 1% penicillin-streptomycin, 1% non-essential amino acids, 0.45μM 1-thioglycerol [2]
Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) Differentiated from embryoid bodies of human ESCs using previously published protocols [26]. Purified using magnetic-activated cell sorting for SIRPA-positive cells [26]. DMEM, 5% fetal bovine serum, 1 mg/mL aminocaproic acid, 50 μg/mL ascorbic acid 2-phosphate, 1 mM sodium pyruvate, 2 mM glutamine, 0.1 mM non-essential amino acids, 0.45 mM 1-thioglycerol [20]
Neonatal rat ventricular myocytes (NRVMs) Enzymatically digested from the ventricles of 2-day-old Sprague Dawley rats. Purified using two pre-plating steps [11]. Low-glucose DMEM, 10% horse serum, 1% chick embryo extract, 1 mg/mL aminocaproic acid, 50 μg/mL ascorbic acid 2-phosphate, 5 U/mL penicillin, 2 μg/mL vitamin B-12 [19]
Mouse induced pluripotent stem cell-derived cardiovascular progenitors (miPSC-CVPs) Differentiated from embryoid bodies of mouse iPSCs using published methods [22]. miPSCs stably transfected with puromycin N-acetyl transferase under the control of a Nkx2–5 cardiac-specific enhancer element to allow for purification of progenitor cells by puromycin selection [22]. 1:1 ratio of DMEM/F12 and Neurobasal base medium, 1x N2 supplement, 1x B-27 supplement, Glutamax-I, non-essential amino acids, β-Mercaptoethanol, 5 mg/mL BSA, Gentamicin, 1 mg/mL aminocaproic acid [22]
Mouse embryonic stem cell-derived cardiomyocytes and cardiovascular progenitors (mESC-CMs and mESC-CVPs) Differentiated from embryoid bodies of mouse ESCs using published protocols [21,27]. mESCs stably transfected with puromycin aminotransferase under the control of a mouse Myh6 promotor or a Nkx2.5 enhancer element to allow for purification by puromycin selection [21]. 1:1 ratio of DMEM/F12 and Neurobasal base medium, 1% L-glutamine, 0.1% gentamicin reagent solution, 1% B-27 supplement, 0.0125 mg/mL insulin, 0.05 mg/mL apo-transferrin, 2.5 ng/mL sodium selenite, 3 ng/mL progesterone, 8 ug/mL putrescine, 0.0334% BSA fraction V, 1 mg/mL aminocaproic acid [21]