Establishment and Application of an in Vitro Cellular Model of Adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells with Serum Injury in aGVHD Mouse.
10.19746/j.cnki.issn.1009-2137.2025.01.038
- Author:
Run-Xiang XU
1
;
Pei-Lin LI
2
;
Jia-Yi TIAN
3
;
Jie TANG
2
;
Bo-Feng YIN
2
;
Fu-Hao YU
1
;
Fei-Yan WANG
2
;
Xiao-Tong LI
2
;
Xiao-Yu ZHANG
2
;
Wen-Rong XIA
4
;
Heng ZHU
2
;
Li DING
1
Author Information
1. Graduate School of Air Force Medical University, Xi'an 710032, Shaanxi Province, China.
2. Beijing Institute of Radiation Medicine, Beijing 100850, China.
3. Department of Hematology, Air Force Medical Center, Air Force Medical University, PLA, Beijing 100142, China.
4. Military Medical College, Beijing 100850, China.
- Publication Type:Journal Article
- Keywords:
hematopoietic stem cell transplantation;
acute graft-versus-host disease;
bone marrow microenvironment;
mesenchymal stem cell;
adipogenesis
- MeSH:
Animals;
Mesenchymal Stem Cells/cytology*;
Mice;
Mice, Inbred BALB C;
Mice, Inbred C57BL;
Adipogenesis;
Female;
Cell Differentiation;
Graft vs Host Disease/blood*;
Bone Marrow Cells/cytology*;
PPAR gamma/metabolism*;
Disease Models, Animal;
CCAAT-Enhancer-Binding Protein-alpha/metabolism*
- From:
Journal of Experimental Hematology
2025;33(1):255-261
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish an in vitro cell model simulating acute graft-versus-host disease (aGVHD) bone marrow microenvironment injury with the advantage of mouse serum of aGVHD model and explore the effect of serum of aGVHD mouse on the adipogenic differentiation ability of mesenchymal stem cells (MSCs).
METHODS:The 6-8-week-old C57BL/6N female mice and BALB/c female mice were used as the donor and recipient mice of the aGVHD model, respectively. Bone marrow transplantation (BMT) mouse model (n=20) was established by being injected with bone marrow cells (1×107 per mouse) from donor mice within 4-6 hours after receiving a lethal dose (8.0 Gy, 72.76 cGy/min) of γ ray general irradiation. A mouse model of aGVHD (n=20) was established by infusing a total of 0.4 ml of a mixture of donor mouse-derived bone marrow cells (1×107 per mouse) and spleen lymphocytes (2×106 per mouse). The blood was removed from the eyeballs and the mouse serum was aspirated on the 7th day after modeling. Bone marrow-derived MSCs were isolated from 1-week-old C57BL/6N male mice and incubated with 2%, 5% and 10% BMT mouse serum and aGVHD mouse serum in the medium, respectively. The effect of serum in the two groups on the in vitro adipogenic differentiation ability of mouse MSCs was detected by Oil Red O staining. The expression levels of related proteins PPARγ and CEBPα were detected by Western blot. The expression differences of key adipogenic transcription factors including PPARγ, CEBPα, FABP4 and LPL were determined by real-time quantitative PCR (RT-qPCR).
RESULTS:An in vitro cell model simulating the damage of bone marrow microenvironment in mice with aGVHD was successfully established. Oil Red O staining showed that the number of orange-red fatty droplets was significantly reduced and the adipogenic differentiation ability of MSC was impaired at aGVHD serum concentration of 10% compared with BMT serum. Western blot experiments showed that adipogenesis-related proteins PPARγ and CEBPα expressed in MSCs were down-regulated. Further RT-qPCR assay showed that the production of PPARγ, CEBPα, FABP4 and LPL, the key transcription factors for adipogenic differentiation of MSC, were significantly reduced.
CONCLUSION:The adipogenic differentiation capacity of MSCs is inhibited by aGVHD mouse serum.
