Ovarian tissue cryopreservation for patients with premature ovary insufficiency caused by cancer treatment: optimal protocol

Li, Y; Ruan, Xiangyan; Liebenthron, Jana; Montag, Markus; Zhou, Qi; Kong, Wei­min; Du, Juan; Jin, Fang; Li, S; Cheng, Jiaojiao; H, Wang; Mueck, Alfred O.

doi:10.1080/13697137.2018.1554644

Cited by 25 publications

(12 citation statements)

References 37 publications

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“…The ovarian tissue was successfully processed and slow-programmatically cryopreserved in ovarian tissue cryobank, and fresh cortex viability and morphology assessment was performed. The methods description was consistent with the previously published article [10,11] . A total of 23 pieces of the ovarian cortex were frozen.…”

Section: Case Presentationmentioning

confidence: 84%

Ovarian Tissue Cryopreservation in a Patient with Breast Cancer During Pregnancy: A Case Report

Cheng¹,

Ruan²,

Du³

et al. 2021

Preprint

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Background: Fertility preservation using ovarian tissue cryopreservation (OTC) in patients with certain diseases especially needing chemo- or radiotherapy is becoming a routine management in various Western countries. Our hospital is the first and until now the only center in China using this method. The question is controversial, if treatment of breast cancer during pregnancy (PrBC) should be similar like for non-pregnant young patients with breast cancer. To our knowledge this worldwide is the first report using OTC as fertility preservation for PrBC. Case presentation: During the 29th week of the pregnancy of a 24-year-old woman needle aspiration cytology of a left breast tumor showed cancer cells. The ultrasound revealed BI-RADS 4a grade. Oncologists recommended the termination of pregnancy. Cesarean section at week 32 was performed, and ovarian tissue samples were collected for OTC to preserve fertility and ovarian endocrine function. Twenty-three ovarian cortex slices were slowly programmed cryopreserved. It is estimated that 13000 follicles have been cryopreserved. Breast nodules and sentinel lymph node biopsy suggested invasive micropapillary carcinoma. Neoadjuvant chemotherapy within 1 week after diagnosis was started. After six courses of neoadjuvant chemotherapy, targeted drug therapy, and Goselin acetate, left mastectomy and left axillary lymph node dissection were performed. In total twenty-three times of radiotherapy, eight trastuzumab targeted therapy, and 17 pertuzumab + trastuzumab double targeted therapy was performed after breast cancer surgery. Until now, more than two years after delivery the ovarian function still is good. We do not see any signs of a negative impact of OTC. The injections of goserelin acetate, every 28 days, are planned to last for the next five years. In addition endocrine therapy with anastrozole, started after the breast cancer surgery, also is scheduled for five years. Conclusion: OTC for fertility preservation in patients with PrBC does not delay breast surgery, radiotherapy, or chemotherapy which is essential for an effective treatment of breast cancer. We assess this method as a promising fertility preservation method, worldwide for the first time now also used in a patient getting breast cancer in pregnancy.

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Section: Case Presentationmentioning

confidence: 84%

Ovarian Tissue Cryopreservation in a Patient with Breast Cancer During Pregnancy: A Case Report

Cheng¹,

Ruan²,

Du³

et al. 2021

Preprint

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“…Follicle viability was evaluated by calcein-AM (Sigma) staining using a protocol published previously (Li et al, 2019). Calcein-AM is a non-fluorescent cell-permeable compound and is converted into fluorescent calcein only in living cells by intracellular esterase in the lysosomes, producing fluorescent green staining.…”

Section: Assessment Of Follicle Viabilitymentioning

confidence: 99%

“…The ROS levels in the fresh and frozen-thawed-cultured ovarian cortex were detected by a spectrofluorimetric method using 2', 7'-dihydro-dichlorofluorescein diacetate (DCHF-DA) assay, as previously described (Li et al, 2019). The TAC assay was conducted based on the Trolox equivalent antioxidant capacity method, as previously described (Li et al, 2019). In brief, this assay assesses the full radical scavenging capacity based on a compound's ability to scavenge the stable 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic diammonium salt radical within 3 min.…”

Section: Assessment Of Ros and Tac In Ovarian Tissuementioning

confidence: 99%

Long-time low-temperature transportation of human ovarian tissue before cryopreservation

Cheng

Ruan

Zhou

et al. 2021

Reproductive BioMedicine Online

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Research question: Can the low-temperature transport time of removed human ovarian tissue be prolonged until cryopreservation? Design: Fresh ovarian cortex from nine premenopausal patients was either slow-programmed frozen immediately or stored at 4℃ for 24h or 48h before slow-freezing. The fresh and frozen-thawed biopsies were evaluated by follicle counting via calcein-staining, histologic analyses via HE, and apoptosis via TUNEL. The fresh cortex was assessed by ROS and TAC assay for oxidative stress 2 2 detection. The frozen-thawed cortex biopsies were also evaluated by qPCR for mRNA expression of BCL-2, BAX, TNFa, HIF-1a, BMP15, and GDF9, western-blot for detection of BCL-2, BMP15, GDF9, and CASPASE3. The frozen-thawed cortex was cultured in-vitro for four-days, AMH and glucose were assessed in the supernatant, and ROS and TAC assay detected cortex's oxidative stress.Results: In the fresh cortex, no significant differences between the three groups. In the frozen-thawed cortex, no significant differences between the three groups regarding follicle viability, TUNEL, mRNA expression of TNFa, HIF-1a, and BMP15. GDF9 mRNA and BAX/BCL-2 were lower and higher at 48h than at 0h, respectively. However, the protein expression of BCL-2, CASPASE3, GDF9, and BMP15 were not different. In the cultured cortex, ROS, TAC, and glucose uptake were not different in the three groups. Conclusion:Ovarian tissue transportation was validated for up to 24h in the practical procedure. This study showed that 4-8℃ transportation for 24h or 48h does not seem to damage the ovarian tissue. However, ovarian tissue transportation above 48h still needs to be further studied for conclusions to be made.

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“…OTC/OTCT is an emerging fertility and endocrine function preservation method, performed to avoid or treat POI [10,11]. Ovarian tissue is frozen before radio-/chemotherapy etc., and later retransplanted [12]. OTC/OTCT is recommended especially if gonadotoxic treatment cannot be delayed and is the exclusive method of preserving fertility in prepubertal girls [11,13].…”

Section: Introductionmentioning

confidence: 99%

“…Within our research we have tried to optimize the protocol for OTC/OTCT [12] and gain new insights for further improvement, like this also is the aim of the present study. Up to now it was not clear how much ovarian tissue can be removed without compromising ovarian function.…”

Section: Introductionmentioning

confidence: 99%

How much total ovarian tissue can be removed without compromising ovarian function? An animal study

Cheng

Ruan

Zhou

et al. 2020

Gynecological Endocrinology

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Purpose Until now, it is not clear how much total ovarian tissue can be removed without affecting ovarian functions. Aim to investigate impact of stepwise removed ovarian tissue on endocrine hormone levels reflecting ovarian function, follicle morphology and weight in rats. MethodsThe study was performed with six groups of animals, each consisting of six ten-week-old Sprague-Dawley rats. A blood test was performed on the day of surgery (baseline) and fortnightly thereafter to assess serum hormones reflecting ovarian function: estradiol (E2), progesterone (P), follicle-stimulating hormone (FSH), anti-Müllerian hormone (AMH), inhibin B (INHB); weight was also assessed. The ovaries obtained from surgical resection and from the rats sacrificed 12 weeks after surgery were stained with hematoxylin-eosin. Surgeries in the groups; [%] of remaining total ovarian tissue: Group 1, non-ovariectomized [100% tissue]. Group 2, half of left ovary excised [75% tissue]. Group 3, left ovary excised [50% tissue]. Group 4, left ovary + half of right ovary excised [25% tissue]. Group 5, left ovary + three quarters of right ovary excised [12.5% tissue]. Group 6, bilateral ovariectomy [0% tissue].Results During the 12 weeks, group 4, 5 and 6 had higher FSH and lower AMH and INHB values compared to the control (group 1) ( p <0.05), but in group 4 production of E2 and P was not significantly different from control ( p >0.05). All ovarian function parameters completely stopped in group 5, i.e. the same as the bilateral ovariectomized rats (group 6). Weight significantly increased in group 6 ( p <0.05). Follicle morphology at baseline and 12 weeks after surgery in groups 2-5 was not significantly different to the control.Conclusions Our animal study confirms that removal of one ovary maintains the remaining ovarian tissue's potency for normal hormone production, as already suggested in clinical studies. However, to our knowledge, we demonstrate here for the first time that even up to 75% of total ovarian tissue can be removed without impact on E2 and P production, which if confirmed in women would mean that hazardous hormone replacement therapy is not required to avoid the negative consequences of hormone depletion.

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Ovarian tissue cryopreservation for patients with premature ovary insufficiency caused by cancer treatment: optimal protocol

Cited by 25 publications

References 37 publications

Ovarian Tissue Cryopreservation in a Patient with Breast Cancer During Pregnancy: A Case Report

Ovarian Tissue Cryopreservation in a Patient with Breast Cancer During Pregnancy: A Case Report

Long-time low-temperature transportation of human ovarian tissue before cryopreservation

How much total ovarian tissue can be removed without compromising ovarian function? An animal study

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