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©The Author(s) 2022.
World J Stem Cells. Aug 26, 2022; 14(8): 633-657
Published online Aug 26, 2022. doi: 10.4252/wjsc.v14.i8.633
Published online Aug 26, 2022. doi: 10.4252/wjsc.v14.i8.633
Table 1 Details of cardiac specific gene primers
No. | Gene primers | Primer sequences (5׳-3׳) | Product size |
1 | GAPDH (F) | GGAAAGCTGTGGCGTGATGG | 414 |
GAPDH (R) | GTAGGCCATGAGGTCCACCA | ||
2 | GATA4 (F) | TCTCACTATGGGCACAGCAG | 245 |
GATA4 (R) | CCGAGCAGGAATTTGAAGAG | ||
3 | cTnT (F) | TTCGACCTGCAGGAAAAGTT | 206 |
cTnT (R) | GTGCCTGGCAAGACCTAGAG | ||
4 | Nkx2.5 (F) | ACCGCCCCTACATTTTATCC | 230 |
Nkx2.5 (R) | GACAGGTACCGCTGTTGCTT | ||
5 | Connexin43 (F) | GCTCCACTCTCGCCTATGTC | 234 |
Connexin43 (R) | GAGTTCATGTCCAGCAGCAA | ||
6 | cMHC (F) | GCAGGTGGATGATCTGGAGG | 116 |
cMHC (R) | CCATGATGCTCTCCTGGGTC | ||
7 | Tbx20 (F) | TGATTCTGGCCCTAGGAAAGTG | 100 |
Tbx20 (R) | TGAGCCTGGGTTCTCCATAAAG | ||
8 | ANF (F) | CCTGCGAAGATCAAGCTGC | 110 |
ANF (R) | GCTCGAGCAGATTTGGCTG | ||
9 | cTnI (F) | GACCTGCGTGGCAAGTTTA | 139 |
cTnI (R) | CCTCCTTCTTCACCTGCTTG | ||
10 | Mef2c (F) | GAGATACCCACAACACACGC | 117 |
Mef2c (R) | GAGTGGAATTCGTTCCGGTG |
Table 2 Degrees of freedom (in vitro experiments)
Experiment | Degrees of freedom (F) |
MTT assay | F3,8 = 404.312 |
Gene expression | |
GATA4 | F3,8 = 14.883 |
cTnT | F3,8 = 15.870 |
cTnI | F3,8 = 34.024 |
Nkx2.5 | F3,8 = 15.901 |
cMHC | F3,8 = 85.276 |
Connexin43 | F3,8 = 19.912 |
ANF | F3,8 = 7.510 |
Tbx20 | F3,8 = 22.877 |
Mef2C | F3,8 = 7.660 |
Protein expression | |
Percent positive cells | |
GATA4 | F3,8 = 26.609 |
cTnT | F3,8 = 46.829 |
Nkx2.5 | F3,8 = 513.556 |
Fluorescence intensity | |
GATA4 | F3,8 = 9.696 |
cTnT | F3,8 = 28.438 |
Nkx2.5 | F3,12 = 59.175 |
Table 3 Echocardiographic parameters
Groups | Duration | LVIDd (cm) | LVIDs (cm) | EF% | FS% | EDV (mL) | ESV (mL) | SV (mL) |
Sham control | 2 wk | 0.4194 | 0.1639 | 93.5889 | 60.9556 | 0.0722 | 0 | 0.0667 |
MI + ve | 0.5156 | 0.3722 | 62.5556 | 28.3056 | 0.1389 | 0.05 | 0.1056 | |
MI + scaffold | 0.5193 | 0.332 | 74.4467 | 37.3067 | 0.1533 | 0.04 | 0.1133 | |
MI + MSC-seeded scaffold | 0.4461 | 0.2611 | 78.8 | 40.9389 | 0.1167 | 0 | 0.0667 | |
MI + zebularine treated MSC-seeded scaffold | 0.5144 | 0.2972 | 80.2111 | 42.1278 | 0.1444 | 0 | 0.1278 | |
Sham control | 4 wk | 0.4705 | 0.195 | 92.7389 | 59.2222 | 0.1222 | 0 | 0.1111 |
MI + ve | 0.5780 | 0.4363 | 57.3306 | 24.9194 | 0.1889 | 0.0833 | 0.1222 | |
MI + scaffold | 0.47 | 0.305 | 73.1056 | 36.3278 | 0.1222 | 0.0333 | 0.0833 | |
MI + MSC-seeded scaffold | 0.5772 | 0.3455 | 78.0056 | 40.5667 | 0.2111 | 0.0389 | 0.1667 | |
MI + zebularine treated MSC-seeded scaffold | 0.5338 | 0.285 | 82.6056 | 45.6611 | 0.1667 | 0 | 0.1222 | |
Sham control | 6 wk | 0.5183 | 0.2333 | 90.3333 | 54.8667 | 0.15 | 0 | 0.1333 |
MI + ve | 0.7066 | 0.5716 | 46.85 | 19.55 | 0.3444 | 0.2 | 0.1611 | |
MI + scaffold | 0.5472 | 0.3622 | 70.0444 | 33.8889 | 0.1667 | 0.0389 | 0.1278 | |
MI + MSC-seeded scaffold | 0.5533 | 0.3288 | 78.05 | 40.8611 | 0.1722 | 0.0333 | 0.1444 | |
MI + zebularine treated MSC-seeded scaffold | 0.5589 | 0.3156 | 81.7028 | 43.7972 | 0.17778 | 0.0167 | 0.1528 |
Table 4 Degrees of freedom (in vivo experiments)
Experiments | Degrees of freedom (F) | ||||
Cardiac function analysis | Parameters | Duration, 2 wk | Duration, 4 wk | Duration, 6 wk | |
LVIDd | F4,25 = 1.936 | F4,25 = 1.299 | F4,25 = 3.594 | ||
LVIDs | F4,25 = 9.060 | F4,25 = 6.944 | F4,25 = 14.008 | ||
%EF | F4,25 = 26.475 | F4,25 = 18.378 | F4,25 = 24.089 | ||
%FS | F4,25 = 35.299 | F4,25 = 24.143 | F4,25 = 26.754 | ||
EDV | F4,25 = 1.571 | F4,25 = 1.059 | F4,25 = 3.601 | ||
ESV | F4,25 = 3.44 | F4,25 = 5.339 | F4,25 = 9.580 | ||
SV | F4,25 = 2.305 | F4,25 = 0.477 | F4,25 = 0.262 | ||
Immunohistochemistry | α-SMA | F4,10 = 31.551 | |||
Cell tracking | Fluorescence intensities of cardiac specific proteins | ||||
cMHC | F = 1.631 | ||||
cTnI | F = 0.882 | ||||
cTnT | F = 2.665 | ||||
Fluorescence intensities of DiI labeled cells | |||||
cMHC | F = 0.398 | ||||
cTnI | F = 0.089 | ||||
cTnT | F = 0.039 |
- Citation: Qazi REM, Khan I, Haneef K, Malick TS, Naeem N, Ahmad W, Salim A, Mohsin S. Combination of mesenchymal stem cells and three-dimensional collagen scaffold preserves ventricular remodeling in rat myocardial infarction model. World J Stem Cells 2022; 14(8): 633-657
- URL: https://www.wjgnet.com/1948-0210/full/v14/i8/633.htm
- DOI: https://dx.doi.org/10.4252/wjsc.v14.i8.633