The Construction of Mesenchymal Stem Cells Carrying Angiopoietin 1 and Its Application in Lung Injury_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

XU Jinfu ¹ ,  QU Jieming ² , SONGLing ³ , HE Lixian ³ , SAI Yin ⁴ , YULong ⁴ , LI Huiping. ¹

Department of Respiratory Medicine, Shanghai Pulmonary Hospital, Tongji University. Shanghai, 200433, ChinaCorresponding Author: QU Jie-Ming, E-mail: jmqu64@ yahoo. com. cn;

Corresponding author：

QU Jieming, Email: jmqu64@yahoo.com.cn

Keywords：

Mesenchymal stem cells; Agiopoietin-1; Vehicle; Gene therapy; Lung injury

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Objective To determine if mesenchymal stem cells ( MSCs) could be reconstructed as a vehicle for angiopoietin-1 ( Ang1) gene therapy in lung injury. Methods MSCs were obtained from adult male inbred mice and cultured to passage four. The cells were identified by fluorescence-activated cell sorting ( FACS) analysis and cell differentiation detection. Lentiviral vectors contained GFP and Ang1 gene were conducted in 293T cells through three plasmids co-transfection method. Then MSCs were transduced with Ang1 gene efficiently through lentiviral vectors. The mRNA expression of Ang1 in MSCs was detected by RT-PCR before and after transfection. Also fluorescence from MSCs was detected by fluorescence microscope every day after transfection. Two hours after LPS inhalation, mice were infused via jugular vein
with normal saline ( NS group) , lentiviral vector carrying Ang1 ( Ang1 group) , lentiviral vector carrying GFP ( MSCs group) , and lentiviral vector carrying Ang1 /GFP ( MSCs-Ang1 group) , respectively. Kaplan-Meier survival analysis was performed to compare the effects of MSCs-Ang1 on survival. And ectogenic MSCs origined lung cells were investigated in receipt mice. Results After passaged and purification,MSCs were confirmed to have the potential of differentiation. The lentiviral vectors carrying Ang1 and GFP were also identified. After transfection, the mRNA expression of Ang1 in MSCs was enhanced. Through the fluorescence microscope,MSCs get the most green fluorescence expression five days after the transfection when MOI was 20. Kaplan-Meier survival analysis showed that MSCs-Ang1 infusion had improved survival rates of lung injury rats compared with the control, but it did not reach statistical significance ( P = 0. 066) . Cells expressing GFP in lung tissues can be observed after MSCs were transplanted in vivo. Conclusions MSCs expressing Ang1 high can be constructed through lentiviral vector transfer, and MSCs-origined cells can be detected in receipt lungs after transplantation. So MSCs may serve as a vehicle for gene therapy in lung injury.

Citation： XU Jinfu,QU Jieming,SONGLing,HE Lixian,SAI Yin,YULong,LI Huiping.. The Construction of Mesenchymal Stem Cells Carrying Angiopoietin 1 and Its Application in Lung Injury. Chinese Journal of Respiratory and Critical Care Medicine, 2009, 09(6): 575-579. doi: Copy

1.	Loebinger MR, Janes SM. Stemcells for lung disease. Chest, 2007 ,132: 279-285.
2.	McCarter SD, Mei SH, Lai PF, et al. Cell-based angiopoietin-1 gene therapy for acute lung injury. Am J Respir Crit Care Med, 2007 ,175: 1014-1026.
3.	Witzenbichler B, Westermann D, Knueppel S, et al. Protective role of angiopoietin-1 in endotoxic shock. Circulation, 2005, 111: 97-105.
4.	Karmpaliotis D, Kosmidou I, Ingenito EP, et al. Angiogenic growth factors in the pathophysiology of a murine model of acute lung injury. Am J Physiol Lung Cell Mol Physiol, 2002, 283: L585 -L595.
5.	郭强, 黄建安, 金钧, 等. 血管生成素-1 对小鼠早期急性肺损伤的保护作用, 中华结核和呼吸杂志, 2007, 30: 926-931.
6.	Peister A, Mellad JA, Larson BL, et al. Adult stem cells from bone marrow ( MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential. Blood, 2004, 103: 1662-1668.
7.	Jeyaseelan S, Chu HW, Young SK, et al. Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury. Infect Immun,2004, 72: 7247-7256.
8.	Rojas M, Xu J, Woods CR, et al. Bone marrow-derived mesenchymal stem cells in repair of the injured lung. Am J Respir Cell Mol Biol, 2005, 33: 145-152.
9.	Snyder JC, Teisanu RM, Stripp BR. Endogenous lung stem cells and contribution to disease. J Pathol, 2009 , 217: 254-264 .
10.	Loebinger MR, Sage EK, Janes SM. Mesenchymal stem cells as vectors for lung disease. Proc Am Thorac Soc, 2008, 5: 711-716.
11.	Gao J, Dennis JE, Muzic RF, et al. The Dynamic in vivo Distribution of Bone Marrow-Derived Mesenchymal Stem Cells after Infusion. Cells Tissues Organs, 2001, 169: 12-20.
12.	Reiser J, Zhang XY, Hemenway CS, et al. Potential of mesenchymal stem cells in gene therapy approaches for inherited and acquired diseases. Expert Opin Biol Ther, 2005, 5: 1571-1584.
13.	Van Damme A, Thorrez L, Ma L, et al. Efficient lentiviral transduction and improved engraftment of human bone marrow mesenchymal cells. StemCells, 2006, 24: 896-907.
14.	Harris RG, Herzog EL, Bruscia EM, et al. Lack of a fusion requirement for development of bone marrow-derived epithelia.Science, 2004, 305: 90-93.

1. Loebinger MR, Janes SM. Stemcells for lung disease. Chest, 2007 ,132: 279-285.
2. McCarter SD, Mei SH, Lai PF, et al. Cell-based angiopoietin-1 gene therapy for acute lung injury. Am J Respir Crit Care Med, 2007 ,175: 1014-1026.
3. Witzenbichler B, Westermann D, Knueppel S, et al. Protective role of angiopoietin-1 in endotoxic shock. Circulation, 2005, 111: 97-105.
4. Karmpaliotis D, Kosmidou I, Ingenito EP, et al. Angiogenic growth factors in the pathophysiology of a murine model of acute lung injury. Am J Physiol Lung Cell Mol Physiol, 2002, 283: L585 -L595.
5. 郭强, 黄建安, 金钧, 等. 血管生成素-1 对小鼠早期急性肺损伤的保护作用, 中华结核和呼吸杂志, 2007, 30: 926-931.
6. Peister A, Mellad JA, Larson BL, et al. Adult stem cells from bone marrow ( MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential. Blood, 2004, 103: 1662-1668.
7. Jeyaseelan S, Chu HW, Young SK, et al. Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury. Infect Immun,2004, 72: 7247-7256.
8. Rojas M, Xu J, Woods CR, et al. Bone marrow-derived mesenchymal stem cells in repair of the injured lung. Am J Respir Cell Mol Biol, 2005, 33: 145-152.
9. Snyder JC, Teisanu RM, Stripp BR. Endogenous lung stem cells and contribution to disease. J Pathol, 2009 , 217: 254-264 .
10. Loebinger MR, Sage EK, Janes SM. Mesenchymal stem cells as vectors for lung disease. Proc Am Thorac Soc, 2008, 5: 711-716.
11. Gao J, Dennis JE, Muzic RF, et al. The Dynamic in vivo Distribution of Bone Marrow-Derived Mesenchymal Stem Cells after Infusion. Cells Tissues Organs, 2001, 169: 12-20.
12. Reiser J, Zhang XY, Hemenway CS, et al. Potential of mesenchymal stem cells in gene therapy approaches for inherited and acquired diseases. Expert Opin Biol Ther, 2005, 5: 1571-1584.
13. Van Damme A, Thorrez L, Ma L, et al. Efficient lentiviral transduction and improved engraftment of human bone marrow mesenchymal cells. StemCells, 2006, 24: 896-907.
14. Harris RG, Herzog EL, Bruscia EM, et al. Lack of a fusion requirement for development of bone marrow-derived epithelia.Science, 2004, 305: 90-93.

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