Hypoxia Rapidly Induces the Expression of Cardiomyogenic Factors in Human Adipose-Derived Adherent Stromal Cells
Abstract
:1. Introduction
2. Experimental Section
2.1. Adipose Tissue Harvesting, Isolation, and Culture of ADASs and ASCs
2.2. Preparation of Normoxic and Hypoxic Conditioned Cells
2.3. Flow Cytometry
2.4. RNA Isolation, Reverse Transcription (RT)-PCR and Quantitive PCR (qPCR) Analysis
2.5. Immunoblot Analysis
2.6. Immunocytochemistry
2.7. Antibody Array
2.8. Statistical Analysis
3. Results
3.1. Characterization of ADASs
3.2. Time-Dependent Differential Regulation of Differentiation-Related Genes of Cell Lineages in ADASs under Hypoxic Conditions
3.3. Time-Dependent Differential Regulation of Cardiomyogenic Proteins in ADASs under Hypoxic Conditions
3.4. Changes in Cardiomyogenic Markers in Dimethyloxaloylglycine -Treated ADASs
3.5. Secretome Analysis of ASCs and ADASs under Hypoxic Conditions
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Genes | Primer Sequence (5′–3′) | |
---|---|---|
HIF1A | F 1 | ACTTGGCAACCTTGGATTGG |
R 2 | GTGCTGAGTAACCACCACTTAT | |
HIF2A | F | CCTGAGACTGTATGGTCAGCTC |
R | AGGACGGAGAGAAGGGAACC | |
PPARG | F | GCAAACCCCTATTCCATGCTG |
R | ACGGAGCTGATCCCAAAGTT | |
LPL | F | CGAGCGCTCCATTCATCTCT |
R | CCAGATTGTTGCAGCGGTTC | |
FABP4 | F | CCTTAGATGGGGGTGTCCTG |
R | AACGTCCCTTGGCTTATGCT | |
GATA4 | F | GCCTCTCGGTGTGACGAGT |
R | TGGTTCCGGAAGCTGATGTAG | |
TBX5 | F | AAGAGTTCCCTCCTCTCCCC |
R | GTCTTGGCCCCGGGAATAAA | |
NKX2.5 | F | CAACATGACCCTGAGTCCCC |
R | TAATCGCCGCCACAAACTCT | |
SOX9 | F | AACAACCCGTCTACACACAGCTCA |
R | TGGGTAATGCGCTTGGATAGGTCA | |
ACAN | F | GTGGTGATGATCTGGCACGAG |
R | CGTTTGTAGGTGGTGGCTGTG | |
COL2A1 | F | TGGTCTTGGTGGAAACTTTGCTGC |
R | AGGTTCACCAGGTTCACCAGGATT | |
COL1A1 | F | CCGGAAACAGACAAGCAACCCAAA |
R | AAAGGAGCAGAAAGGGCAGCATTG | |
RUNX2 | F | AAGGGTCCACTCTGGCTTTG |
R | CTAGGCGCATTTCAGGTGCT | |
OCN | F | TCACACTCCTCGCCCTATT |
R | TGAAAGCCGATGTGGTCAG | |
ALPL | F | GACCCTTGACCCCCACAAT |
R | CGCCTCGTACTGCATGTCCCCT | |
TNNT2 | F | GACAGAGCGGAAAAGTGGGA |
R | CACAGCTCCTTGGCCTTCTC | |
MYH6 | F | ACCAACCTGTCCAAGTTCCG |
MYH7 | F | GACACACTTGAGTAGCCCAGG |
R | CTTCTAGCCGCTCCTTCTCTG | |
Internal control | ||
GAPDH | F | CATGGGTGTGAACCATGAGA |
R | GGTCATGAGTCCTTCCACGA |
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Choi, J.-W.; Moon, H.; Jung, S.E.; Lim, S.; Lee, S.; Kim, I.-K.; Lee, H.-B.; Lee, J.; Song, B.-W.; Kim, S.W.; et al. Hypoxia Rapidly Induces the Expression of Cardiomyogenic Factors in Human Adipose-Derived Adherent Stromal Cells. J. Clin. Med. 2019, 8, 1231. https://doi.org/10.3390/jcm8081231
Choi J-W, Moon H, Jung SE, Lim S, Lee S, Kim I-K, Lee H-B, Lee J, Song B-W, Kim SW, et al. Hypoxia Rapidly Induces the Expression of Cardiomyogenic Factors in Human Adipose-Derived Adherent Stromal Cells. Journal of Clinical Medicine. 2019; 8(8):1231. https://doi.org/10.3390/jcm8081231
Chicago/Turabian StyleChoi, Jung-Won, Hanbyeol Moon, Seung Eun Jung, Soyeon Lim, Seahyoung Lee, Il-Kwon Kim, Hoon-Bum Lee, Jiyun Lee, Byeong-Wook Song, Sang Woo Kim, and et al. 2019. "Hypoxia Rapidly Induces the Expression of Cardiomyogenic Factors in Human Adipose-Derived Adherent Stromal Cells" Journal of Clinical Medicine 8, no. 8: 1231. https://doi.org/10.3390/jcm8081231