Table 2.
Makers for characterizing myocytes and non-myocytes derived from PSC cultures
| Cell type | Surface marker expression by flow cytometry | Gene marker expression by PCR, bulk and/or sRNA-seq | Protein marker expression by immunohistochemistry | Electrophysiological properties | References |
|---|---|---|---|---|---|
| Myocytes | |||||
| Pan/ventricular | cTnT+, SIRPA+, NKX2-5+, VCAM+, MLC2V+, MYL2+, CD77+CD200−, HEY2+ | cTnT, NKX2-5, MLC2V, MYL2, IRX4, MEF2C, MYH6, MYH7, GJA1, PDLIM1, VTN, LPL, HEY2 | cTnT, MLC2V, MYL2, NKX2-5, MYH7, α-actinin, VCAM | Fast upstroke velocity (> 10 V/s), longer AP duration, plateau phase | [15••, 72, 87, 104, 107•, 108, 110, 115•, 116–119] |
| Immature ventricular | - | KDR, ISL, GATA4, TNNI1; Churko et al. [121•] noted the following genes were enriched in early-stage cardiomyocytes, i.e. day 14: VCAM1, APOE, NPPA, NPPB, MYH6, COL2A1, HMGA1, S100A10, ATF4, RBFOX2, HMGA2, CLDN6 | - | Slow upstroke velocity (< 10 V/s), slow beating rate (50 bpm) | [86, 104, 118, 119] |
| Mature ventricular | CD36+SIRPA+LDLR+ | MYH7, HEY2; Churko et al. [121•] noted the following genes were enriched in day 45 cardiomyocytes: HEY2, FHL2, CAV1, SERPINI1, MYL2, A2M, LGALS3BP, TMEM173, SYNE2, NFIA, HOPX; Funakoshi et al. [113•] classified mature cells with the following highly expressed genes: FAO: CD36, FABP3, ACSL1; Mitochondrial: CKMT2, COX6A2; Muscle function: DES, MYBPC3, ACTN2 and further classified cell types as non-proliferating, non-stressed (CD36, LDLR), stressed (ATF5, TRIB3), proliferating (MKI167, FOXM1) and cardiac fibroblasts (FN1, COL3A1) | - | - | [104, 110, 118] |
| Atrial | MLC2A+ | ACTA2, ATP2A2, CACNA1D, GJA5 (CX40), HEY1, KCNA5, KCNK3, KCNJ3, MLC2A, MYH6, MYLK, MYL7, NR2F1, NR2F2, NPPA, SLN, TBX5, RBP1 | MLC2A | Triangular morphology | [77, 87, 110, 115•, 116–119] |
| Pacemaker | - | Transcription factor: SHOX2, ISL1, TBX3, TBX5, TBX18; ion channel: HCN1, HCN4, KCNJ3, SCN5A | TBX3 | Fast spontaneous firing rates, slow maximum upstroke velocity, small AP amplitudes, short AP durations; faster beating rates | [104, 115•, 117, 119] |
| SAN pacemaker | - | ISL1, SHOX2, TBX3, TBX18 | SHOX2 | - | [87, 104, 115•] |
| AVN pacemaker | - | MSX2, TBX2 | - | - | [115•] |
| Non-myocytes | |||||
| Pluripotency markers | OCT4+, SOX2+, TRA160+, SSEA-4+ | POU5F1, DNMT3B, NANOG | OCT4, NANOG, TRA-1–80, SSEA-4 | N/A | [15••, 82, 87, 104] |
| Endoderm | - | SOX17, FOXA2 | - | N/A | [104] |
| Mesoderm | Flk-1+PdgfR-α+, CD235a+ (ventricular mesoderm), RALDH2+ (atrial mesoderm), CD13+ (early cardiac mesoderm) | EOMES, T, MESP1, MIXL1 | - | N/A | [66, 87, 104, 118–120] |
| Fibroblast | CD90+ | CD90 | P4HB, TE-7 | N/A | [15••, 87, 104, 108, 119] |
| Epithelial | - | - | PanCK | N/A | [15••] |
| Endothelial | VE-cad+, PECAM+, CD31+, CD34+ | CDH5, CD34, PECAM, TAL1 | vWF, CD31 | N/A | [15••, 72, 104, 107•, 108, 115•, 121•] |
| Vascular smooth muscle | CNN1+ | ACTA2, CNN1, MYH11, TAGLIN2 | αSMA | N/A | [71, 86, 104, 107•] |