Effects of Human Amnion–Derived Mesenchymal Stem Cell (hAD-MSC) Transplantation In Situ on Primary Ovarian Insufficiency in SD Rats

Abstract

Human amnion–derived mesenchymal stem cell (hAD-MSC) transplantation can repair ovarian injury and improve ovarian function in rats with chemotherapy-induced primary ovarian insufficiency (POI). However, ensuring that stem cells home to the ovary to improve their effects on ovarian injury is challenging. This research aimed to directly inject ovarian tissue with hAD-MSCs and improve the homing of stem cells to the ovary. The animals were divided into POI, hAD-MSC (tail vein) treatment, hAD-MSC (in situ) treatment, and control groups. POI rat models were established by intraperitoneal injection of cyclophosphamide (CTX) and busulfan (BUS). The hAD-MSCs isolated from the amnion were injected into the tail vein or ovary of POI rats. The estrous cycle, serum sex hormone levels, follicle counts, ovarian pathological changes, and proteome of the ovaries were evaluated. hAD-MSCs were successfully isolated and cultured from the amnion. Both hAD-MSC (tail vein) and hAD-MSC (in situ) transplantation increased body weight, improved the AMH levels and follicle numbers, and reduced reproductive organ injuries in POI rats. Transplantation of hAD-MSCs (in situ) upregulated 24 proteins and downregulated 4 proteins. Both hAD-MSC (tail vein) and hAD-MSC (in situ) transplantations can repair ovarian injury and improve ovarian function in rats with chemotherapy-induced POI. The paracrine proteome of hAD-MSCs in the ovarian microenvironment can protect against chemotherapy-induced damage by reducing apoptosis and promoting angiogenesis, cell proliferation, and gene expression.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Data Availability

The datasets used and/or analyzed during the current study are available from the corresponding author on a reasonable request.

Abbreviations

POI:

primary ovarian insufficiency

CTX:

cyclophosphamide

BUS:

busulfan

CTX:

cyclophosphamide

DAPI:

2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride

EDTA:

ethylenediaminetetraacetic acid

ELISA:

enzyme-linked immunosorbent assay

FGF:

fibroblast growth factor

FSH:

follicle-stimulating hormone

GC:

granulosa cell

H&E:

hematoxylin and eosin

hAD-MSC:

human amnion–derived mesenchymal stem cell

IGF:

insulin-like growth factor

IL:

interleukin

MSC:

mesenchymal stem cell

OCT:

optimal cutting temperature

POF:

premature ovarian failure

POI:

primary ovarian insufficiency

SD:

Sprague-Dawley

VEGF:

vascular endothelial growth factor

FBS:

fetal bovine serum

E2:

estradiol

ANOVA:

analysis of variance

ERK:

extracellular signal–regulated kinase

JNK:

c-Jun N-terminal kinase

Akt:

protein kinase B

PI3K:

phosphatidylinositol-3-kinase

MKK:

MAPK kinase

PF4:

platelet factor 4

MAPK:

mitogen-activated protein kinase

TNF:

tumor necrosis factor

MMP:

matrix metalloproteinase

References

  1. 1.

    Siegel RL, Miller KD, Jemal A. Cancer statistics, 2018. CA Cancer J Clin. 2018;68(1):7–30.

    Article  Google Scholar 

  2. 2.

    De Vos M, Devroey P, Fauser BC. Primary ovarian insufficiency. Lancet. 2010;376(9744):911–21.

    Article  Google Scholar 

  3. 3.

    Levine J, Canada A, Stern CJ. Fertility preservation in adolescents and young adults with cancer. J Clin Oncol. 2010;28(32):4831–41.

    Article  Google Scholar 

  4. 4.

    Tonorezos ES, Hudson MM, Edgar AB, Kremer LC, Sklar CA, Wallace WH, et al. Screening and management of adverse endocrine outcomes in adult survivors of childhood and adolescent cancer. Lancet Diabetes Endocrinol. 2015;3(7):545–55.

    Article  Google Scholar 

  5. 5.

    Overbeek A, van den Berg MH, van Leeuwen FE, Kaspers GJ, Lambalk CB, van Dulmen-den Broeder E. Chemotherapy-related late adverse effects on ovarian function in female survivors of childhood and young adult cancer: a systematic review. Cancer Treat Rev. 2017;53:10–24.

    CAS  Article  Google Scholar 

  6. 6.

    Atsma F, Bartelink ML, Grobbee DE, van der Schouw YT. Postmenopausal status and early menopause as independent risk factors for cardiovascular disease: a meta-analysis. Menopause. 2006;13(2):265–79.

    Article  Google Scholar 

  7. 7.

    Bove R, Secor E, Chibnik LB, Barnes LL, Schneider JA, Bennett DA, et al. Age at surgical menopause influences cognitive decline and Alzheimer pathology in older women. Neurology. 2014;82(3):222–9.

    Article  Google Scholar 

  8. 8.

    Tao XY, Zuo AZ, Wang JQ, Tao FB. Effect of primary ovarian insufficiency and early natural menopause on mortality: a meta-analysis. Climacteric. 2016;19(1):27–36.

    Article  Google Scholar 

  9. 9.

    Ling L, Feng X, Wei T, Wang Y, Wang Y, Zhang W, et al. Effects of low-intensity pulsed ultrasound (LIPUS)-pretreated human amnion-derived mesenchymal stem cell (hAD-MSC) transplantation on primary ovarian insufficiency in rats. Stem Cell Res Ther. 2017;8(1):283.

    Article  Google Scholar 

  10. 10.

    Luo Q, Yin N, Zhang L, Yuan W, Zhao W, Luan X, et al. Role of SDF-1/CXCR4 and cytokines in the development of ovary injury in chemotherapy drug induced premature ovarian failure mice. Life Sci. 2017;179:103–9.

    CAS  Article  Google Scholar 

  11. 11.

    Wei Wei, Ximei Wu, Li Y. Experimental methodology of pharmacology, 4th edition. 2010:1698.

  12. 12.

    Wang J, WU S, Shang H. Observation of estrous cycle in sexually mature female SD rats. Contemp Med. 2013;19(28):25–6.

    Google Scholar 

  13. 13.

    Wang Z, Wang Y, Yang T, Li J, Yang X. Study of the reparative effects of menstrual-derived stem cells on premature ovarian failure in mice. Stem Cell Res Ther. 2017;8(1):11.

    Article  Google Scholar 

  14. 14.

    Pedersen T, Peters H. Proposal for a classification of oocytes and follicles in the mouse ovary. J Reprod Fertil. 1968;17(3):555–7.

    CAS  Article  Google Scholar 

  15. 15.

    Faddy MJ. Follicle dynamics during ovarian ageing. Mol Cell Endocrinol. 2000;163(1–2):43–8.

    CAS  Article  Google Scholar 

  16. 16.

    European Society for Human R, Embryology Guideline Group on POI, Webber L, et al. ESHRE guideline: management of women with premature ovarian insufficiency. Hum Reprod. 2016;31(5):926–37.

    Article  Google Scholar 

  17. 17.

    Jensen AK, Rechnitzer C, Macklon KT, Ifversen MR, Birkebæk N, Clausen N, et al. Cryopreservation of ovarian tissue for fertility preservation in a large cohort of young girls: focus on pubertal development. Hum Reprod. 2017;32(1):154–64.

    CAS  PubMed  Google Scholar 

  18. 18.

    Su J, Ding L, Cheng J, Yang J, Li X, Yan G, et al. Transplantation of adipose-derived stem cells combined with collagen scaffolds restores ovarian function in a rat model of premature ovarian insufficiency. Hum Reprod. 2016;31(5):1075–86.

    CAS  Article  Google Scholar 

  19. 19.

    Levine JM, Kelvin JF, Quinn GP, Gracia CR. Infertility in reproductive-age female cancer survivors. Cancer. 2015;121(10):1532–9.

    Article  Google Scholar 

  20. 20.

    Sullivan SD, Sarrel PM, Nelson LM. Hormone replacement therapy in young women with primary ovarian insufficiency and early menopause. Fertil Steril. 2016;106(7):1588–99.

    CAS  Article  Google Scholar 

  21. 21.

    Demeestere I, Brice P, Peccatori FA, Kentos A, Dupuis J, Zachee P, et al. No evidence for the benefit of gonadotropin-releasing hormone agonist in preserving ovarian function and fertility in lymphoma survivors treated with chemotherapy: final long-term report of a prospective randomized trial. J Clin Oncol. 2016;34(22):2568–74.

    CAS  Article  Google Scholar 

  22. 22.

    Herraiz S, Buigues A, Diaz-Garcia C, et al. Fertility rescue and ovarian follicle growth promotion by bone marrow stem cell infusion. Fertil Steril. 2018;109(5):908–18 e902.

    Article  Google Scholar 

  23. 23.

    Ling L, Feng X, Wei T, Wang Y, Wang Y, Wang Z, et al. Human amnion-derived mesenchymal stem cell (hAD-MSC) transplantation improves ovarian function in rats with premature ovarian insufficiency (POI) at least partly through a paracrine mechanism. Stem Cell Res Ther. 2019;10(1):46.

    CAS  Article  Google Scholar 

  24. 24.

    Ling L, Wei T, He L, et al. Low-intensity pulsed ultrasound activates ERK1/2 and PI3K-Akt signalling pathways and promotes the proliferation of human amnion-derived mesenchymal stem cells. Cell Prolif. 2017, 50(6):e12383.

  25. 25.

    Sipp D, Robey PG, Turner L. Clear up this stem-cell mess. Nature. 2018;561(7724):455–7.

    CAS  Article  Google Scholar 

  26. 26.

    Meirow D, Biederman H, Anderson RA, Wallace WH. Toxicity of chemotherapy and radiation on female reproduction. Clin Obstet Gynecol. 2010;53(4):727–39.

    Article  Google Scholar 

  27. 27.

    Bukovsky A. Cell commitment by asymmetric division and immune system involvement. Prog Mol Subcell Biol. 2007;45:179–204.

    CAS  Article  Google Scholar 

  28. 28.

    Ye H, Zheng T, Li W, Li X, Fu X, Huang Y, et al. Ovarian stem cell nests in reproduction and ovarian aging. Cell Physiol Biochem. 2017;43(5):1917–25.

    CAS  Article  Google Scholar 

  29. 29.

    White YA, Woods DC, Takai Y, Ishihara O, Seki H, Tilly JL. Oocyte formation by mitotically active germ cells purified from ovaries of reproductive-age women. Nat Med. 2012;18(3):413–21.

    CAS  Article  Google Scholar 

  30. 30.

    Lin H. The stem-cell niche theory: lessons from flies. Nat Rev Genet. 2002;3(12):931–40.

    CAS  Article  Google Scholar 

  31. 31.

    Niikura Y, Niikura T, Tilly JL. Aged mouse ovaries possess rare premeiotic germ cells that can generate oocytes following transplantation into a young host environment. Aging (Albany NY). 2009;1(12):971–8.

    CAS  Article  Google Scholar 

  32. 32.

    Massasa E, Costa XS, Taylor HS. Failure of the stem cell niche rather than loss of oocyte stem cells in the aging ovary. Aging (Albany NY). 2010;2(1):1–2.

    Article  Google Scholar 

  33. 33.

    Bukovsky A, Caudle MR. Immunoregulation of follicular renewal, selection, POF, and menopause in vivo, vs. neo-oogenesis in vitro, POF and ovarian infertility treatment, and a clinical trial. Reprod Biol Endocrinol. 2012;10:97.

    Article  Google Scholar 

  34. 34.

    Winkler IG, Pettit AR, Raggatt LJ, Jacobsen RN, Forristal CE, Barbier V, et al. Hematopoietic stem cell mobilizing agents G-CSF, cyclophosphamide or AMD3100 have distinct mechanisms of action on bone marrow HSC niches and bone formation. Leukemia. 2012;26(7):1594–601.

    CAS  Article  Google Scholar 

  35. 35.

    Sun L, Akiyama K, Zhang H, Yamaza T, Hou Y, Zhao S, et al. Mesenchymal stem cell transplantation reverses multiorgan dysfunction in systemic lupus erythematosus mice and humans. Stem Cells. 2009;27(6):1421–32.

    CAS  Article  Google Scholar 

  36. 36.

    Lai D, Wang F, Yao X, Zhang Q, Wu X, Xiang C. Human endometrial mesenchymal stem cells restore ovarian function through improving the renewal of germline stem cells in a mouse model of premature ovarian failure. J Transl Med. 2015;13:155.

    Article  Google Scholar 

  37. 37.

    Zhang D, Liu Y, Zhang Z, Lv P, Liu Y, Li J, et al. Basonuclin 1 deficiency is a cause of primary ovarian insufficiency. Hum Mol Genet. 2018;27(21):3787–800.

    CAS  Article  Google Scholar 

  38. 38.

    Tuppi M, Kehrloesser S, Coutandin DW, Rossi V, Luh LM, Strubel A, et al. Oocyte DNA damage quality control requires consecutive interplay of CHK2 and CK1 to activate p63. Nat Struct Mol Biol. 2018;25(3):261–9.

    CAS  Article  Google Scholar 

  39. 39.

    Rehnitz J, Alcoba DD, Brum IS, Hinderhofer K, Youness B, Strowitzki T, et al. FMR1 and AKT/mTOR signalling pathways: potential functional interactions controlling folliculogenesis in human granulosa cells. Reprod BioMed Online. 2017;35(5):485–93.

    CAS  Article  Google Scholar 

  40. 40.

    Yin N, Wang Y, Lu X, Liu R, Zhang L, Zhao W, et al. hPMSC transplantation restoring ovarian function in premature ovarian failure mice is associated with change of Th17/Tc17 and Th17/Treg cell ratios through the PI3K/Akt signal pathway. Stem Cell Res Ther. 2018;9(1):37.

    Article  Google Scholar 

  41. 41.

    Feng P, Li P, Tan J. Human menstrual blood-derived stromal cells promote recovery of premature ovarian insufficiency via regulating the ECM-dependent FAK/AKT signaling. Stem Cell Rev. 2019, 15(2):241–255.

  42. 42.

    Kawamura K, Kawamura N, Hsueh AJ. Activation of dormant follicles: a new treatment for premature ovarian failure? Curr Opin Obstet Gynecol. 2016;28(3):217–22.

    Article  Google Scholar 

  43. 43.

    Zhou Y, Qin Y, Qin Y, Xu B, Guo T, Ke H, et al. Wdr62 is involved in meiotic initiation via activating JNK signaling and associated with POI in humans. PLoS Genet. 2018;14(8):e1007463.

    Article  Google Scholar 

  44. 44.

    Kim J, Perez AS, Claflin J, David A, Zhou H, Shikanov A. Synthetic hydrogel supports the function and regeneration of artificial ovarian tissue in mice. NPJ Regen Med. 2016;1.16010.

Download references

Funding

This study was supported by the National Natural Science Foundation of China (81671415).

Author information

Affiliations

Authors

Contributions

Conceived and designed the experiments: XF and ZX; literature search: XF and LL; performed the experiments: XF, LL, WZ, XL, YL, and DT; analyzed the data: XF and LL; helped perform the analysis with constructive discussions: XF, LL, YW, and ZX; drafted the manuscript: XF and LL. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Zhengai Xiong.

Ethics declarations

This research was in accordance with the Helsinki Declaration and approved by the Ethics Committee of the Second Affiliated Hospital of Chongqing Medical University.

Competing Interests

The authors declared that they have no conflict of interest.

Ethics Approval and Consent to Participate

The protocol was approved by the Committee on the Ethics of Animal Experiments of Chongqing Medical University (Permit Number: 2016-044).

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Feng, X., Ling, L., Zhang, W. et al. Effects of Human Amnion–Derived Mesenchymal Stem Cell (hAD-MSC) Transplantation In Situ on Primary Ovarian Insufficiency in SD Rats. Reprod. Sci. 27, 1502–1512 (2020). https://doi.org/10.1007/s43032-020-00147-0

Download citation

Keywords

  • hAD-MSCs
  • Primary ovarian insufficiency
  • Cyclophosphamide
  • Busulfan
  • Proteome