Cesarean section has no impact on bone marrow mesenchymal stem cell homing in pregnant rats

doi:10.3969/j.issn.2095-4344.0885

Abstract

Abstract:

BACKGROUND: Long-term complications of cesarean section include placenta praevia, placenta accreta, cesarean scar pregnancy and uterine rupture. These life-threatening complications in pregnant women maybe result from the defects of endometrium and uterine smooth muscle as well as poorly formed decidua in the scar of cesarean section. Mesenchymal stem cells have the function of repairing tissue injuries, and the amount of cells homing to the site of injury may affect the effect of tissue repair.
OBJECTIVE: To explore the distribution and homing of bone marrow mesenchymal stem cells from male rats into rat models of cesarean section.
METHODS: Bone marrow mesenchymal stem cells isolated from male rats in vitro were cultured and identified. Female Wistar rats were randomized into two groups: cesarean section group and control group. Rats in the cesarean section group were given intravenous administration of bone marrow mesenchymal stem cells from male rats via the tail vein on day 21 after cesarean section, and non-operative rats in the control groups were given the same amount of bone marrow mesenchymal stem cells from male rats after natural delivery. Rats in the two groups were sacrificed on days 7 and 28 after cell injection. The distribution of bone marrow mesenchymal stem cells from male rats in tissues (including heart, lungs, livers, kidneys, and uterine scar) was detected by measurement of the SRY mRNA level using SPY-PCR.
RESULTS AND CONCLUSION: In the cesarean section group, the SRY gene was most abundant in the lung, followed by the liver and the kidney on day 7 after injection, although the distribution of SRY gene in the heart and uterine scar was low; on day 28 after injection, the levels of SRY gene in the lung, liver and kidney decreased (P < 0.05), but had no significant changes in the heart and uterine scar (P > 0.05). In the control group, the distribution of SRY gene was similar to that in the cesarean section group on both days 7 and 28 after injection, and the levels of SRY gene in the heart and uterus were low. These findings reveal that allogeneic bone marrow mesenchymal stem cells implanted mainly distribute in tissues with abundant blood flow, including lungs, livers and kidneys. And the cell number decreases gradually over time. Since the amount of implanted cells in the heart and uterus is very low, the change with time is not obvious. Cesarean section injury has no impact on the distribution of bone marrow mesenchymal stem cells in pregnant rats and there is of course no increase in the homing and colonization of bone marrow mesenchymal stem cells in a cesarean scar.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Mesenchymal Stem Cell Transplantation, Cesarean Section, Genes, sry, Stem Cells

CLC Number:

R394.2

Sha Wen-qiong, She Rui-lian, Wang Shuo-shi, Wang Qian, Xu Man, Shi Wei, Li Lan. Cesarean section has no impact on bone marrow mesenchymal stem cell homing in pregnant rats[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(17): 2644-2649.

Figures/Tables 1

2.1 大鼠BMSCs的培养和鉴定大鼠BMSCs呈长梭形贴壁生长，并具有较好的增殖能力。在成骨培养基和成脂培养基中体外三四周的诱导，大鼠BMSCs可向骨细胞及脂类细胞进一步分化，可分化为成脂细胞及成骨细胞(图1)。流式细胞仪检测结果提示：BMSCs的CD29、CD44、CD73、CD90、CD105和CD166均呈阳性表达，表达效率分别为96.2%，87.9%，94.7%，89.8%，93.7%和92.9%，BMSCs的CD14、CD34和CD45均呈阴性表达，表达效率分别为0.23%，0.34%和0.54% 。 2.2 实验动物数量分析入组的24只大鼠在处死前没有死亡，全部进入结果分析。 2.3 SRY-PCR结果 SRY基因是雄性的性别决定基因。此次研究中体外培养雄性大鼠BMSCs，经尾静脉注射入雌性大鼠体内，雌性大鼠处死后，检测组织中SRY基因含量，间接反映了组织中雄性大鼠BMSCs的存留量。两组大鼠经尾静脉注射雄性大鼠BMSCs后，通过RT-PCR技术检测大鼠各个组织(心、肺、肝、肾、以及子宫瘢痕)中SRY基因的相对含量，从而评价BMSCs的分布水平。注射后7 d，剖宫产手术组大鼠SRY基因在肺组织中含量最高(0.073 1±0.018 4)，其次在肝组织中(0.012 6± 0.002 8)，在肾组织中也分布一定比例(0.005 3±0.002 7)，而在心脏(0.000 6±0.000 1)和子宫瘢痕(0.001 0±0.000 3)处的SRY基因含量较少。注射后28 d，各组织中SRY基因的相对含量与7 d的检测结果略有不同，主要体现在肺(0.039 6±0.008 4)、肝(0.009 5±0.003 3)和肾(0.003 9± 0.001 6)中的相对含量均有一定程度的下降(P < 0.05)，而在心脏(0.000 8±0.000 3)和子宫瘢痕(0.001 0±0.000 2)处，SRY基因的相对含量未见显著变化(P > 0.05)。见图2。在非手术对照组(图3)，孕鼠注射雄性大鼠BMSCs 7 d后，SRY基因仍然在肺组织中含量最高(0.070 5±0.023 4)，其次在肝组织中含量较多(0.013 4±0.002 3)，在肾组织中也分布一定比例(0.006 1±0.005 5)，在心脏(0.0006± 0.0002)和子宫(0.001 1±0.000 4)处的SRY基因含量较少；注射28 d肺(0.038 4±0.020 4)、肝(0.009 1±0.002 7)和肾(0.004 0±0.001 9)中的相对含量均有一定程度的下降，而在心脏(0.000 7±0.000 3)和子宫(0.000 9±0.000 3)处，SRY的相对含量未见显著变化，总体来讲，与剖宫产手术组的检测结果相似。"

References

[1] Lumbiganon P, Laopaiboon M, Gülmezoglu AM,et al.Method of delivery and pregnancy outcomes in Asia: the WHO global survey on maternal and perinatal health 2007-08.Lancet. 2010;375(9713):490-499.

[2] 程蔚蔚,朱关珍.剖宫产的近期和远期并发症[J].中国实用妇科与产科杂志, 2003,19(7):442-444.

[3] Roberge S, Demers S, Berghella V,et al.Impact of single- vs double-layer closure on adverse outcomes and uterine scar defect: a systematic review and metaanalysis.Am J Obstet Gynecol. 2014;211(5):453-460.

[4] Fukuchi Y, Nakajima H, Sugiyama D, et al. Human placenta-derived cells have mesenchymal stem/progenitor cell potential.Stem Cells. 2004;22(5):649-658.

[5] Pittenger MF, Mackay AM, Beck SC et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284(5411):143-147.

[6] Baksh D, Yao R, Tuan RS. Comparison of proliferative and multilineage differentiation potential of human mesenchymal stem cells derived from umbilical cord and bone marrow. Stem Cells. 2007;25(6):1384-1392.

[7] Campagnoli C, Roberts IA, Kumar S,et al. Identification of mesenchymal stem/progenitor cells in human first trimester fetal blood, liver, and bone marrow. Blood. 2001;98(8): 2396-2402.

[8] Kinnaird T, Stabile E, Burnett MS,et al.Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms.Circ Res. 2004; 94(5):678-685.

[9] Williams AR, Hare JM. Mesenchymal Stem Cells: Biology, Pathophysiology, Translational Findings, and Therapeutic Implications for Cardiac Disease. Circ Res. 2011;109(8): 923-940

[10] Haleem AM, Singergy AA, Sabry D,et al.The Clinical Use of Human Culture-Expanded Autologous Bone Marrow Mesenchymal Stem Cells Transplanted on Platelet-Rich Fibrin Glue in the Treatment of Articular Cartilage Defects: A Pilot Study and Preliminary Results.Cartilage. 2010;1(4): 253-261.

[11] Tang Y, Cai B, Yuan F, et al. Melatonin pretreatment improves the survival and function of transplanted mesenchymal stem cells after focal cerebral ischemia. Cell Transplant.2013; 23(10):1279-1291.

[12] Jin Y, Bouyer J, Shumsky JS,et al. Transplantation of neural progenitor cells in chronic spinal cord injury.Neuroscience. 2016;320:69-82.

[13] Mathiasen AB，Qayyum AA, Jorgensen E，et al．Bone marrow derived mesenchymal stromal cell treatment in patients with severe ischaemic heart failure: a randomized placebo- controlled trial(MSC-HF trial)．Eur Heart J.2015; 36(27): 1744-1753．

[14] Jayaraman P, Nathan P, Vasanthan P, et al. Stem cells conditioned medium: a new approach to skin wound healing management. Cell Biol Int.2013;37(10):1122-1128.

[15] Patel DM, Shah J, Srivastava AS. Therapeutic potential of mesenchymal stem cells in regenerative medicine. Stem Cells Int.2013;2013:496218.

[16] Shabbir A, Cox A, Rodriguez-Menocal L, et al. Mesenchymal Stem Cell Exosomes Induce Proliferation and Migration of Normal and Chronic Wound Fibroblasts, and Enhance Angiogenesis In Vitro. Stem Cells Dev. 2016;24(14): 1635-1647.

[17] Wang Y, Chen X, Cao W, et al. Plasticity of mesenchymalstem cells in immunomodulation: pathological and therapeuticimplications. Nat Immunol. 2014;15(11): 1009-1016.

[18] Hashemian SJ, Kouhnavard M, Nasli-Esfahani E.Mesenchymal stem cells: rising concerns over theirapplication in treatment of type one diabetes mellitus. J Diabetes Res. 2015;1:1-19

[19] 高杰清,谢宗燕,母义明,等.脐带间充质干细胞在2型糖尿病大鼠中的疗效及作用机制[J].中华内分泌代谢杂志, 2017,33(1): 62-67.

[20] Saito Y, Shimada M, Utsunomiya T,et al.Homing effect of adipose-derived stem cells to the injured liver: the shift of stromal cell-derived factor 1 expressions.J Hepatobiliary Pancreat Sci. 2014;21(12):873-880.

[21] 邓蓉蓉,谢伊旻,谢林.间充质干细胞归巢的研究与进展[J].中国组织工程研究,2016,10(19):2879-2888.

[22] Ringe J, Strassburg S, Neumann K, et al. Towards insitu tissue repair: human mesenchymal stem cellsexpress chemokine receptors CXCR1, CXCR2 andCCR2, and migrate upon stimulation with CXCL8 butnot CCL2. J Cell Biochem. 2007;101(1):135-146.

[23] Weeks S,Kulkarni A,Smith H,et al. The effects of chemokine, adhesion and extracellular matrix molecules on binding of mesenchymal stromal cells to poly( l -lactic acid). Cytotherapy. 2012;14(9):1080-1088.

[24] 程斌,方驰华,范应方,等.骨髓间充质干细胞提取方法的改良[J].中国组织工程研究与临床康复, 2008,12(47):9293-9296.

[25] Karp JM, Leng Teo GS. Mesenchymal stem cellhoming: the devil is in the details. Cell Stem Cell.2009;4(3):206-216.

[26] Cheng K, Rai P, Plagov A,et al.Transplantation of bone marrow-derived MSCs improves cisplatinum-induced renal injury through paracrine mechanisms.Exp Mol Pathol. 2013; 94(3):466-473.

[27] Gao J, Dennis JE, Muzic RF, et al. The dynamic in vivodistribution of bone marrow-drived mesenehymal stem cellsafter infusion. Cells Tissues Organs. 2001;169(1):12-20.

[28] Liu J, Wang H, Li J.Inflammation and Inflammatory Cells in Myocardial Infarction and Reperfusion Injury: A Double-Edged Sword.Clin Med Insights Cardiol. 2016;10:79-84.

[29] Halabian R, Tehrani HA, Jahanian-Najafabadi A, et al. Lipocalin-2-mediated upregulation of various antioxidants and growth factors protects bone marrow-derived mesenchymal stem cells against unfavorable microenvironments.Cell Stress Chaperones. 2013;18(6):785-800.

[30] Siao CJ,Lorentz CU,Kermani P,et al．ProNGF，a cytokine induced after myocardial infarction in humans，targets pericytesto promote microvascular damage and activation.J Exp Med. 2012;209(12):2291-2305．

[31] 张静,魏峰,吴忠东,等.同种异体骨髓间充质于细胞在大鼠体内心肌缺血微环境中的分化及存活[J].安徽医科大学学报, 2014, 49(3):300-303．

[32] LV di Bonzo LV, Ferrero I, Cravanzola C, et al. Human mesenchymal stem cells as a two-edged sword in hepatic regenerative medicine: engraftment andhepatocyte differentiation versus profibrogenic potential. Gut. 2008; 57(2):223-231.

[33] 何秀华,李东良,江军,等.移植途径对大鼠骨髓间充质干细胞归巢及肝功能恢复的影响[J].中华细胞与干细胞杂志:电子版, 2011, 1(1): 57-62.