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Isolation of CD146+ Resident Lung Mesenchymal Stromal Cells from Rat Lungs
Published on: June 17, 2016
[Marrow-derived mesenchymal stem cells protect the lung injury caused by exposure to high oxygen: experiment with
Zhao-fang Tian1, Yu-hong Li, Xue-mei Fu
1Department of Neonatology, Huai'an First Hospital Affiliated to Nanjing Medical University, Huai'an 223300, China.
Objective:
To investigate the influence of marrow-derived mesenchymal stem cells (MSCs) on the lung injury caused by exposure to high oxygen.
Methods:
MSCs were extracted from the femurs of Sprague-Dawley (SD) rat, cultured, amplified, and labeled with 5-bromo-2'-deoxy-uridine (BrdU). Thirty-two 3-day-old SD rats were randomly divided into 4 equal groups: Group A, exposed to high oxygen (95%) for 7 days and then injected intra-peritoneally with 50 microl phosphate buffered solution (PBS) containing 5 x 10(4) MSCs; Group B, exposed to ordinary air for 7 days and then injected intra-peritoneally with PBS containing 5 x 10(4) MSCs; Group C, exposed to high oxygen (95%) for 7 days and then injected intra-peritoneally with PBS without MSC; and Group D, exposed to ordinary air for 7 days and then injected intra-peritoneally with PBS without MSC. 72 h later, when the rats were 13 days old they were killed Bronchoalveolar lavage fluid (BALF) was obtained to count the numbers of white blood cells and neutrophils. Lung homogenate was made. ELISA was used to examine the contents of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta. Radial alveolar count (RAC) of the lungs was conducted under light microscope. Double-staining immunocytochemistry was used to detect the expression of surfactant protein A (SP-A) and BrdU.
Results:
(1) The RAC values of Groups A and C were (11.0 +/- 1.0) and (9.6 +/- 0.7) cells/10 fields, both significantly lower than those of Group B and D [(13.9 +/- 1.1) and (14.0 +/- 1.1) cells/10 fields respectively, F = 40.41, P = 0.000]. The RAC of Group A was significantly higher than that of Group C (P < 0.05). (2) The white blood cell numbers in BALF of Groups A and C were (0.85 +/- 0.21) and (1.44 +/- 0.69) x 10(5)/ml respectively, both significantly higher than those of Groups B and D [(0.18 +/- 0.10) and (0.21 +/- 0.06) x 10(5)/ml respectively, F = 21.52, P = 0.000]. The neutrophil numbers of Groups A and C were (0.32 +/- 0.12) and (0.73 +/- 0.35) x 10(5)/ml respectively, both significantly higher than those of Groups B and D [(0.06 +/- 0.02) and (0.07 +/- 0.02) x 10(5)/ml respectively, F = 22.91, P = 0.000]. (3) The TNF-alpha levels in the homogenate of Groups A and C were (173 +/- 20) and (224 +/- 42) pg/m respectively, both significantly higher than those of Group B and D [(34 +/- 4) and (35 +/- 4) pg/ml respectively, F = 138.1, P = 0.000]. The IL-1beta levels in the homogenate of Groups A and C were (530 +/- 74) and (948 +/- 82) pg/ml respectively, both significantly higher than those of Group B and D [(210 +/- 33) and (216 +/- 30) pg/ml respectively, F = 247.4, P = 0.000]. (4) BrdU-positive cells and SP-A-positive cells were seen in both Groups A and B, a few double-stained cells were found in Group A, but only SP-A-positive cells, and none BrdU positive cells and double-stained cells were seen in Groups C and D.
Conclusion:
MSC administration has protective effect on the ling injury caused by exposure to high oxygen via different mechanisms.
Insights
Marrow-derived mesenchymal stem cells (MSCs) show a protective effect against high oxygen-induced lung injury in rats. MSC treatment improved lung structure and reduced inflammation, suggesting therapeutic potential.
Area of Science:
- Regenerative Medicine
- Pulmonary Medicine
- Cell Biology
Background:
- High oxygen exposure can cause significant lung injury, particularly in premature infants.
- Marrow-derived mesenchymal stem cells (MSCs) are being investigated for their therapeutic potential in various conditions.
Purpose of the Study:
- To investigate the protective effects of MSCs on lung injury induced by high oxygen exposure.
- To explore the underlying mechanisms of MSC-mediated protection.
Main Methods:
- Mesenchymal stem cells (MSCs) were extracted, cultured, and labeled in Sprague-Dawley rats.
- Rats were exposed to high oxygen or normal air and treated with MSCs or placebo.
- Lung injury was assessed by bronchoalveolar lavage fluid analysis, inflammatory cytokine levels (TNF-alpha, IL-1beta), radial alveolar count, and cell marker expression (BrdU, SP-A).
Main Results:
- MSC treatment in high oxygen conditions (Group A) showed improved radial alveolar counts compared to controls (Group C).
- MSC administration reduced white blood cell and neutrophil infiltration in the lungs.
- Inflammatory markers (TNF-alpha, IL-1beta) were significantly lower in MSC-treated groups.
- Evidence of MSC engraftment and surfactant protein A expression was observed.
Conclusions:
- Marrow-derived mesenchymal stem cell administration demonstrates a protective effect against high oxygen-induced lung injury.
- The protective mechanisms involve structural improvements, reduced inflammation, and potential cell differentiation or integration.
