Cargando…

Effects of hypoxia-preconditioned HucMSCs on neovascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice

BACKGROUND: The massive loss of follicles in the early stage of ovarian tissue transplantation is considered a significant restriction to the efficacy of ovarian tissue cryopreservation (OTC) and transplantation (OT). The use of mesenchymal stem cells (MSCs) before transplantation of ovarian fragmen...

Descripción completa

Detalles Bibliográficos
Autores principales: Cheng, Jiaojiao, Ruan, Xiangyan, Li, Yanglu, Du, Juan, Jin, Fengyu, Gu, Muqing, Zhou, Qi, Xu, Xin, Yang, Yu, Wang, Husheng, Mueck, Alfred Otto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9476266/
https://www.ncbi.nlm.nih.gov/pubmed/36104746
http://dx.doi.org/10.1186/s13287-022-03167-6
_version_ 1784790099951616000
author Cheng, Jiaojiao
Ruan, Xiangyan
Li, Yanglu
Du, Juan
Jin, Fengyu
Gu, Muqing
Zhou, Qi
Xu, Xin
Yang, Yu
Wang, Husheng
Mueck, Alfred Otto
author_facet Cheng, Jiaojiao
Ruan, Xiangyan
Li, Yanglu
Du, Juan
Jin, Fengyu
Gu, Muqing
Zhou, Qi
Xu, Xin
Yang, Yu
Wang, Husheng
Mueck, Alfred Otto
author_sort Cheng, Jiaojiao
collection PubMed
description BACKGROUND: The massive loss of follicles in the early stage of ovarian tissue transplantation is considered a significant restriction to the efficacy of ovarian tissue cryopreservation (OTC) and transplantation (OT). The use of mesenchymal stem cells (MSCs) before transplantation of ovarian fragments shortened the hypoxic period and boosted neovascularization. Hypoxia-preconditioned MSCs can enhance the potential of angiogenesis. Can hypoxia-preconditioned human umbilical cord mesenchymal stem cell (HucMSCs) and ovarian tissue co-xenotransplantation improve more neovascularization and subsequently more follicle survival in human ovarian tissue? METHODS: Frozen-thawed cortical pieces from 4 patients were transplanted into the bilateral renal capsule of immune-deficient nude mice without HucMSCs or normoxia/hypoxia-preconditioned HucMSCs. Sixty-four mice were randomly distributed into 4 groups. In each group, the mice were euthanized for blood and/or graft retrieval on post-transplantation days 3 (n = 8) and 7 (n = 8), respectively. Non-grafted frozen-thawed ovarian fragment was taken for non-grafted control. Grafts were histologically processed and analysed for follicle density and atretic follicles by HE, neovascularization by CD34 and CD31 immunohistochemical staining, primordial follicle growth by Ki67 staining, and apoptosis of stromal cell and follicles by immunofluorescence using TUNEL. The ROS and TAC levels of grafted and non-grafted tissue were assessed. We evaluated the protein expression of HIF1α, VEGFA, pAkt, Akt, and GDF9 in grafted and non-grafted ovarian tissue. E2, Prog, AMH, and FSH levels in the plasma of mice were measured after 3 and 7 days of OT. RESULTS: Hypoxia-preconditioned HucMSCs positively protect the grafted ovarian tissue by significantly decreasing the apoptosis and increasing higher expression of CD31, CD34, and VEGFA for earlier angiogenesis. They are crucial to preserving the resting primordial follicle pool by modulation of follicle death. CONCLUSION: This is the first study to demonstrate that co-transplantation of hypoxia-preconditioned HucMSC with ovarian tissue improved earlier vascularization of ovarian grafts in the early post-grafting period, which correlates with increased follicle survival and reduced apoptosis. The HIF1α/VEGFA signal pathways may play an important role in elucidating the mechanisms of action of hypoxia-preconditioned HucMSCs with regard to OT and clinical implementation.
format Online
Article
Text
id pubmed-9476266
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-94762662022-09-16 Effects of hypoxia-preconditioned HucMSCs on neovascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice Cheng, Jiaojiao Ruan, Xiangyan Li, Yanglu Du, Juan Jin, Fengyu Gu, Muqing Zhou, Qi Xu, Xin Yang, Yu Wang, Husheng Mueck, Alfred Otto Stem Cell Res Ther Research BACKGROUND: The massive loss of follicles in the early stage of ovarian tissue transplantation is considered a significant restriction to the efficacy of ovarian tissue cryopreservation (OTC) and transplantation (OT). The use of mesenchymal stem cells (MSCs) before transplantation of ovarian fragments shortened the hypoxic period and boosted neovascularization. Hypoxia-preconditioned MSCs can enhance the potential of angiogenesis. Can hypoxia-preconditioned human umbilical cord mesenchymal stem cell (HucMSCs) and ovarian tissue co-xenotransplantation improve more neovascularization and subsequently more follicle survival in human ovarian tissue? METHODS: Frozen-thawed cortical pieces from 4 patients were transplanted into the bilateral renal capsule of immune-deficient nude mice without HucMSCs or normoxia/hypoxia-preconditioned HucMSCs. Sixty-four mice were randomly distributed into 4 groups. In each group, the mice were euthanized for blood and/or graft retrieval on post-transplantation days 3 (n = 8) and 7 (n = 8), respectively. Non-grafted frozen-thawed ovarian fragment was taken for non-grafted control. Grafts were histologically processed and analysed for follicle density and atretic follicles by HE, neovascularization by CD34 and CD31 immunohistochemical staining, primordial follicle growth by Ki67 staining, and apoptosis of stromal cell and follicles by immunofluorescence using TUNEL. The ROS and TAC levels of grafted and non-grafted tissue were assessed. We evaluated the protein expression of HIF1α, VEGFA, pAkt, Akt, and GDF9 in grafted and non-grafted ovarian tissue. E2, Prog, AMH, and FSH levels in the plasma of mice were measured after 3 and 7 days of OT. RESULTS: Hypoxia-preconditioned HucMSCs positively protect the grafted ovarian tissue by significantly decreasing the apoptosis and increasing higher expression of CD31, CD34, and VEGFA for earlier angiogenesis. They are crucial to preserving the resting primordial follicle pool by modulation of follicle death. CONCLUSION: This is the first study to demonstrate that co-transplantation of hypoxia-preconditioned HucMSC with ovarian tissue improved earlier vascularization of ovarian grafts in the early post-grafting period, which correlates with increased follicle survival and reduced apoptosis. The HIF1α/VEGFA signal pathways may play an important role in elucidating the mechanisms of action of hypoxia-preconditioned HucMSCs with regard to OT and clinical implementation. BioMed Central 2022-09-14 /pmc/articles/PMC9476266/ /pubmed/36104746 http://dx.doi.org/10.1186/s13287-022-03167-6 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Cheng, Jiaojiao
Ruan, Xiangyan
Li, Yanglu
Du, Juan
Jin, Fengyu
Gu, Muqing
Zhou, Qi
Xu, Xin
Yang, Yu
Wang, Husheng
Mueck, Alfred Otto
Effects of hypoxia-preconditioned HucMSCs on neovascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice
title Effects of hypoxia-preconditioned HucMSCs on neovascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice
title_full Effects of hypoxia-preconditioned HucMSCs on neovascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice
title_fullStr Effects of hypoxia-preconditioned HucMSCs on neovascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice
title_full_unstemmed Effects of hypoxia-preconditioned HucMSCs on neovascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice
title_short Effects of hypoxia-preconditioned HucMSCs on neovascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice
title_sort effects of hypoxia-preconditioned hucmscs on neovascularization and follicle survival in frozen/thawed human ovarian cortex transplanted to immunodeficient mice
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9476266/
https://www.ncbi.nlm.nih.gov/pubmed/36104746
http://dx.doi.org/10.1186/s13287-022-03167-6
work_keys_str_mv AT chengjiaojiao effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT ruanxiangyan effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT liyanglu effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT dujuan effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT jinfengyu effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT gumuqing effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT zhouqi effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT xuxin effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT yangyu effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT wanghusheng effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice
AT mueckalfredotto effectsofhypoxiapreconditionedhucmscsonneovascularizationandfolliclesurvivalinfrozenthawedhumanovariancortextransplantedtoimmunodeficientmice