Dimethyl sulfoxide maintains structure and function of cryopreserved equine endometrial explants

Thompson, Riley E.; Johnson, Aime K.; Prado, Tulio M; Premanandan, Christopher; Brown, Megan E.; Whitlock, Brian K; Pukazhenthi, Budhan S.

doi:10.1016/j.cryobiol.2019.10.006

Cited by 9 publications

(3 citation statements)

References 28 publications

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“…Conversely, when functional assessment is required, then balanced and optimal cellular performance must take precedence over structure. This indicates an interesting, and valuable, halfway house for cryopreservation where success can be measured in terms of either the structural integrity or function of recovered material [37,38].…”

Section: The Cryobiology Of Scale Up 21 Practical Challengesmentioning

confidence: 99%

Scaling up Cryopreservation from Cell Suspensions to Tissues: Challenges and Successes

Kilbride¹,

Meneghel²,

Chawda³

et al. 2023

Biomedical Engineering

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This chapter covers the key physical, biological and practical challenges encountered when developing cryopreservation protocols for larger biological structures and examines areas where cryopreservation has been successful in scaling to larger structures. Results from techniques being used in attempts to overcome these challenges are reviewed together with the indicators for future development that arise from them. The scale-up of cryopreservation to tissues with diverse functions and cell types makes the control of freezing and thawing more challenging. Technology may—or may not—be available depending on the size of the material involved. To meet the challenge there must be innovation in technology, techniques and understanding of damage-limiting strategies. Diversity of cell structure, size, shape and expected function means a similarly diverse response to any imposed cryopreservation conditions and interaction with ice crystals. The increasing diffusion distances involved, and diversity of permeability properties, will affect solutes, solvents, heat and cryoprotectant (CPA) transfer and so add to the diversity of response. Constructing a single protocol for cryopreservation of a larger sample (organoids to whole organs) becomes a formidable challenge.

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Section: The Cryobiology Of Scale Up 21 Practical Challengesmentioning

confidence: 99%

Scaling up Cryopreservation from Cell Suspensions to Tissues: Challenges and Successes

Kilbride¹,

Meneghel²,

Chawda³

et al. 2023

Biomedical Engineering

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“…Fetal tissues were transported and isolated as above (see Organoid Culture). Cells were scraped from the basement membrane in Handling Medium [HM; MEM with Earle's Salts, 25 mM HEPES, 100 U/mL Penicillin, 0.1 mg/mL Streptomycin, 0.1 mM Pyruvate, 2 mM Glutamax, 5% Bovine Serum Albumin] [39,40] and then centrifuged at 600× g for 6 min. The resulting pellet was resuspended in equine chorionic girdle monolayer medium [Monolayer medium: DMEM/F12 without phenol red with 2.5 mM L-glutamine, 100 U/mL penicillin-streptomycin, 10% (v/v) FBS] [34] and plated at 250 mL per well in a 48-well plate.…”

Section: Monolayer Cell Culturementioning

confidence: 99%

Production of Mare Chorionic Girdle Organoids That Secrete Equine Chorionic Gonadotropin

Thompson

Meyers

Palmer

et al. 2023

IJMS

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The equine chorionic girdle is comprised of specialized invasive trophoblast cells that begin formation approximately 25 days after ovulation (day 0) and invade the endometrium to become endometrial cups. These specialized trophoblast cells transition from uninucleate to differentiated binucleate trophoblast cells that secrete the glycoprotein hormone equine chorionic gonadotropin (eCG; formerly known as pregnant mare serum gonadotropin or PMSG). This eCG has LH-like activity in the horse but variable LH- and FSH-like activity in other species and has been utilized for these properties both in vivo and in vitro. To produce eCG commercially, large volumes of whole blood must be collected from pregnant mares, which negatively impacts equine welfare due to repeated blood collections and the birth of an unwanted foal. Attempts to produce eCG in vitro using long-term culture of chorionic girdle explants have not been successful beyond 180 days, with peak eCG production at 30 days of culture. Organoids are three-dimensional cell clusters that self-organize and can remain genetically and phenotypically stable throughout long-term culture (i.e., months). Human trophoblast organoids have been reported to successfully produce human chorionic gonadotropin (hCG) and proliferate long-term (>1 year). The objective of this study was to evaluate whether organoids derived from equine chorionic girdle maintain physiological functionality. Here we show generation of chorionic girdle organoids for the first time and demonstrate in vitro production of eCG for up to 6 weeks in culture. Therefore, equine chorionic girdle organoids provide a physiologically representative 3D in vitro model for chorionic girdle development of early equine pregnancy.

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“…For example, mare endometrial explants immersed in culture media degenerate significantly over a 48-h culture period [ 3 ]. An alternative equine endometrial explant model that utilized a gas–fluid interface by placing explants on agarose blocks partially immersed in culture media ( Figure 1 C) facilitated maintenance of tissue for 5 days in culture, but often formed a central core of necrosis [ 4 , 5 ]. However, long-term cultures (weeks to months) are necessary to fully recapitulate physiological reproductive processes over one or multiple estrous cycles or to evaluate pathologies with an insidious onset.…”

Section: In Vitro Models Used To Study Reproductive Physiologymentioning

confidence: 99%

The Roles of Extracellular Vesicles and Organoid Models in Female Reproductive Physiology

Thompson

Bouma

Hollinshead

2022

IJMS

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Culture model systems that can recapitulate the anatomy and physiology of reproductive organs, such as three-dimensional (3D) organoid culture systems, limit the cost and welfare concerns associated with a research animal colony and provide alternative approaches to study specific processes in humans and animals. These 3D models facilitate a greater understanding of the physiological role of individual cell types and their interactions than can be accomplished with traditional monolayer culture systems. Furthermore, 3D culture systems allow for the examination of specific cellular, molecular, or hormonal interactions, without confounding factors that occur with in vivo models, and provide a powerful approach to study physiological and pathological reproductive conditions. The goal of this paper is to review and compare organoid culture systems to other in vitro cell culture models, currently used to study female reproductive physiology, with an emphasis on the role of extracellular vesicle interactions. The critical role of extracellular vesicles for intercellular communication in physiological processes, including reproduction, has been well documented, and an overview of the roles of extracellular vesicles in organoid systems will be provided. Finally, we will propose future directions for understanding the role of extracellular vesicles in normal and pathological conditions of reproductive organs, utilizing 3D organoid culture systems.

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Dimethyl sulfoxide maintains structure and function of cryopreserved equine endometrial explants

Cited by 9 publications

References 28 publications

Scaling up Cryopreservation from Cell Suspensions to Tissues: Challenges and Successes

Scaling up Cryopreservation from Cell Suspensions to Tissues: Challenges and Successes

Production of Mare Chorionic Girdle Organoids That Secrete Equine Chorionic Gonadotropin

The Roles of Extracellular Vesicles and Organoid Models in Female Reproductive Physiology

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