Human umbilical cord blood mononuclear cell transplantation for treatment of radioactive premature ovarian failure in nude mice

doi:10.3969/j.issn.2095-4344.2016.36.012

Abstract

Abstract:

BACKGROUND: Stem cell transplantation, in recent years, has become a preferred treatment for premature ovarian failure. Umbilical cord blood mononuclear cells containing a large number of mesenchymal stem cells, immature stem/progenitor cells, and endothelial progenitor cells can be used as an important source of stem cell transplantation.
OBJECTIVE: To study the therapeutic efficacy of human umbilical cord blood mononuclear cell transplantation on radioactive premature ovarian failure in nude mice.
METHODS: 120 female BALB/C nude mice were randomly divided into four groups: blank control group without any intervention; model group, intravenous transplantation group, and in situ transplantation group exposed to 60Co γ rays, 0.5 Gry per day, for 30 days. After 30 days, premature ovarian failure models were made in the latter three groups. Then, nude mice in the latter three groups were given bilateral ovary injection of 10 µL DMEM, tail vein injection of 10 µL human umbilical cord blood mononuclear cells (1×10¹³/L), and bilateral ovary injection of 10 µL human umbilical cord blood mononuclear cells (1×10¹³/L), respectively. Thirty days after cell transplantation, serum levels of estradiol, follicle hormone, luteinizing hormone, inhibin B and vascular endothelial growth factor were detected, cell apoptosis in the ovary tissue and cell survival were observed pathologically.
RESULTS AND CONCLUSION: After modeling, the serum levels of estradiol, inhibin B and vascular endothelial growth factor were significantly reduced (P < 0.01), while follicle hormone and luteinizing hormone levels increased (P < 0.01). After transplantation, these indexes were all improved in the in situ transplantation group (P < 0.01), and reduced follicle hormone and luteinizing hormone levels were visible in the intravenous transplantation group (P< 0.05,P < 0.01). Compared with the model group, in situ transplantation and intravenous injection of human umbilical cord blood mononuclear cells could effectively reduce cell apoptosis in the ovary tissue (P < 0.05), and transplanted cells were able to survive in the ovary of nude mice. All these findings show human umbilical cord blood mononuclear cell transplantation do have curative effects on premature ovarian failure in nude mice through the inhibition of apoptosis and the regulation of hormone secretion.

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: Stem Cells, Stem Cell Transplantation, Ovarian Diseases, Tissue Engineering

CLC Number:

R394.2

Wang Yi-feng, Song Wen-guang, Liu Shu-xia . Human umbilical cord blood mononuclear cell transplantation for treatment of radioactive premature ovarian failure in nude mice[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(36): 5398-5404.

Figures/Tables 4

2.1 实验动物数量分析 120只大鼠均进入结果分析。 2.2 各组卵巢功能指标检测结果由方差分析结果可知，空白对照组、模型组、静脉移植组、原位移植组之间比较差异极其显著(F=61.99，P < 0.0001)。与空白对照组比较，模型组雌二醇、抑制素B、血管内皮生长因子水平降低(P < 0.01)，卵泡激素、黄体生成素水平升高(P < 0.01)。与模型组比较，原位移植组雌二醇、抑制素B、血管内皮生长因子水平升高(P < 0.01)，卵泡激素、黄体生成素水平降低(P < 0.01)；静脉移植组卵泡激素、黄体生成素水平降低(P < 0.05，P < 0.01)，见表1。提示人脐血单个核细胞原位移植对卵巢早衰有一定的治疗意义，静脉移植人脐血单个核细胞对卵巢早衰也有一定意义，但与原位移植相比存在一定差异。 2.3 各组Tunel染色结果由图1中观察可知，绿色荧光光点为凋亡细胞，在各图之中，空白对照组凋亡细胞颗粒极少；模型组荧光光点最多，表明模型组中凋亡细胞颗粒最多；与模型组相比，原位移植组、静脉移植组荧光光点较少，原位移植组凋亡细胞颗粒较静脉移植组多。以颗粒细胞数为指标，进行统计学分析发现，各组与模型组相比都具有统计学意义(表2)，提示静脉及原位移植人脐血单个核细胞可有效减少卵巢中的颗粒细胞的凋亡，改善裸鼠卵巢功能。 2.4 移植后人脐血单个核细胞存活情况利用免疫组化方法在裸鼠卵巢中寻找被BrdU标记的人脐血单个核细胞，发现其在裸鼠卵巢内可以定居并存活，其结果见图2。模型组裸鼠卵巢组织切片染色呈阴性，提示模型组卵巢组织中无人脐血单个核细胞；尾静脉注射组裸鼠卵巢组织切片可见经标记后的人脐血单个核细胞，表明人脐血单个核细胞在移植后可在裸鼠卵巢中存活。"

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