TABLE 1.
Description of basic properties of our synergistic three-phase differentiation protocol and protocols developed by Patsch et al. (2015) (nicknamed high VEGF + forskolin protocol in our article) and Sahara et al. (2014) (nicknamed DAPT protocol in our article).
| Protocol | Duration of differentiation | Primitive streak and mesoderm differentiated by separate media (precision of differentiation) | Derived cells tested for standard surface markers | Cytokines used in each stage of differentiation: primitive streak = PS, mesoderm = M, endothelium = E | Media used in each stage of differentiation: primitive streak = PS, mesoderm = M, endothelium = E | Number of hPSC lines tested (robustness) | Average percentage of successfully differentiated cells (efficiency) | Precise single cell seeding density/media volumes defined (reproducibility and standardization) |
| Our synergistic three phase protocol (Farkas et al.) | 5 days | Yes | CD31, CD34, CD144, KDR | PS: CP21R7, BMP4, FGF2 M: BMP4, FGF2 E: high concentration VEGF165, DAPT, Forskolin | PS: STEMdiff APEL2 M: STEMdiff APEL2 E: STEMdiff APEL2 | 7 | 85–94% for CD31, CD34, CD144, KDR | Yes – 400 000 cells per pm35 well*/yes – 1 ml per pm35 well in stage 1, 2 ml per well in stage 2 and three |
| Patsch et al., 2015 (original article results) | 5 days | No | CD31, CD144, vWF | M: BMP4, CP21R7 E: high concentration VEGF165, Forskolin | M: N2B27 + Neurobasal medium E: StemPro34 | 4 | 70.1 % CD144 | 370000–470000/No |
| Patsch et al., 2015 (our test results) | 5 days | No | CD31, CD34, CD144, KDR | M: BMP4, CP21R7 E: high concentration VEGF165, Forskolin | M: N2B27 + Neurobasal medium E: StemPro34 | 2 | 49–61% for CD31, CD34, CD144, KDR | Yes – 400 000 cells per pm35 well */yes 3 ml per pm35 well in each stage, no refreshment of medium – most efficient method we tested |
| Sahara et al., 2014 (original article results) | 5 days | No | CD31, CD34, CD144, KDR | M: BMP4, CP21R7 E: VEGF165, DAPT | M: N2B27 + Neurobasal medium + DMEM-F12 E: StemPro34 | 4 | 50% CD31 + CD144 | No/No |
| Sahara et al., 2014 (our test results) | 5 days | No | CD31, CD34, CD144, KDR | M: BMP4, CP21R7 E:VEGF165, DAPT | M: N2B27 + Neurobasal medium + DMEM-F12 E: StemPro34 | 2 | 44–54% for CD31, CD34, CD144, KDR | Yes – 400 000 cells per pm35 well* /yes 3 ml per pm35 well in each stage, no refreshment of medium – most efficient method we tested |
Table includes relevant properties such as duration, precision, robustness, efficiency, reproducibility and standardization of differentiation, tested surface markers, cytokines, and media used in each stage of differentiation. Table includes information from both original articles written by their respective authors and our tests separately. *pm35 dish 10 cm2.