This patient's CPE resolved when intrapleural alteplase was used as an adjunct to chest tube drainage and antibiotics. Controlled trials need to be conducted to investigate fully the efficacy, dosing, and safety of intrapleural alteplase in the treatment of patients with CPE and empyema.

This study investigated how follicle health and development in human ovarian tissue cryopreserved for fertility preservation varied between patients before and after six days of in vitro culture. Ovarian tissue from 12 patients (9-25 years) was used. In 3 patients, a 1hr neutral red (NR) incubation was used to identify tissues with viable follicles. Tissues were fixed, sectioned, and follicles staged and graded for health. Inter-patient differences were observed in the non-cultured tissue in the number of both healthy follicles (p=0.005) and growing follicles (p=0.005). After culture there was significant variation in the number of transitional, primary, and secondary follicles between patients (p<0.001).Asymmetric primary follicles with a single complete layer of granulosa cells plus two or more additional partial layers were 5.5 times more likely to be observed in cultured compared to non-cultured tissue (p=0.0063). Non-cultured (p=0.0125) and cultured (p<0.001) tissue selected using NR had more healthy follicles compared to tissue not selected using NR. Non-cultured and cultured tissue selected using NR had more healthy follicles compared to tissue not selected using NR (p=0.0125; p<0.001). We demonstrate that inter-patient variation exists in the health and development of follicles before and after culture. Culture systems need to be optimized to support cryopreserved ovarian tissue and these findings should prompt researchers to consider patient variation when evaluating culture systems.

In vitro follicle growth is a potential fertility preservation method for patients for whom current methods are contraindicated. Currently this method has only been successful using fresh ovarian tissue. Since many patients who may benefit from this treatment currently have cryopreserved ovarian tissue in storage, optimising in vitro follicle growth (IVG) for cryopreserved-thawed tissue is critical. This study sought to improve the first step of IVG by comparing different short-term culture systems for cryopreserved-thawed human ovarian tissue, in order to yield a higher number of healthy multilayer follicles. We compared two commonly used culture media (αMEM and McCoy’s 5A), and three plate conditions (300 µL, 1 mL on a polycarbonate membrane and 1 mL in a gas-permeable plate) on the health and development of follicles after six days of culture. A total of 5,797 follicles from three post-pubertal patients (aged 21.3 ± 2.3 years) were analysed across six different culture conditions and non-cultured control. All culture systems supported follicle development and there was no difference in developmental progression between the different conditions tested. Differences in follicle morphology were evident with follicles cultured in low volume conditions having significantly greater odds of being graded as morphologically normal compared to other conditions. Furthermore, culture in a low volume of αMEM resulted in the highest proportion of morphologically normal primary and multilayer follicles (23.8% compared to 6.3-19.9% depending on condition). We therefore recommend culture of cryopreserved human ovarian tissue in a low volume of αMEM to support follicle health and development.

Background. Ovarian tissue cryopreservation (OTC) is important for fertility preservation and conservation. Delay in OTC can occur for transport or workflow management, yet little is understood about the effect of this on the tissue. Objective. To determine if a delay of 24-48 h to OTC affects primordial follicle (PF) health. Methods. Ovaries (n = 6 sheep) were processed immediately or after storage at 4°C (24 h, 48 h). Tissue was fixed fresh, after cryopreservation or 10-day xenotransplantation. Morphological assessment of follicle health and development was performed. Findings. A total of 1,541 follicles were analysed. A 24 h processing delay did not impact PF health in fresh or cryopreserved tissue. In fresh tissue, a 48 h delay had an adverse effect on PF health (OR = 2.47, 95% CI 1.29-4.71). Interestingly, a 48 h delay resulted in cryopreserved tissue being less likely to be graded as abnormal compared to control (OR = 0.56, 95% CI 0.36-0.87). There was no difference in PF health or development across groups following xenotransplantation. Conclusion. Ovarian tissue can be stored for up to 48 h prior to cryopreservation with no net impact on PF morphology which is indicative of health.

AbstractBackgroundOvarian tissue cryopreservation (OTC) is important for fertility preservation and conservation. Delay in OTC may be required for transport or workflow management, however little is understood about the effect of processing delay on the tissue.ObjectiveTo determine whether a delay of 24-48 hours to OTC affects primordial follicle (PF) health.MethodsOvaries (n=6 sheep) were processed immediately or after storage at 4°C (24h, 48h). Tissue was fixed fresh, after cryopreservation or 10-day xenotransplantation. Morphological assessment of follicle health and development was performed.FindingsA total of 1541 follicles were analysed. A 24h processing delay did not impact PF health in fresh or cryopreserved tissue. In fresh tissue a 48h delay had an adverse effect on follicle health (OR=2.47, 95% CI 1.29-4.71). Interestingly, a 48h delay resulted in cryopreserved tissue being less likely to be graded as unhealthy compared to control (OR=0.56, 95% CI 0.36-0.87). There was no difference in PF health or development across groups following xenotransplantation.ConclusionOvarian tissue can be stored for up to 48 hours prior to cryopreservation with no net impact on PF health.