Embryo rescue is a vital technique in cucurbit breeding and propagation, addressing challenges such as embryo abortion, poor seed viability, and incompatibility barriers. This method involves the excision of immature embryos from seeds followed by their in vitro culture on a nutrient medium, providing an environment conducive to their growth and development. In cucurbits, embryo rescue has been extensively utilized to overcome barriers to hybridization, enabling the production of interspecific and intergeneric hybrids with desired traits. Various factors, including genotype, developmental stage of embryos, and culture conditions, influence the success of embryo rescue in cucurbits. Optimal nutrient formulations, growth regulators, and culture techniques are critical for promoting embryo germination, shoot elongation, and subsequent plantlet establishment. Additionally, embryo rescue facilitates the recovery of valuable genetic material from wild and exotic cucurbit species, expanding genetic diversity and developing novel cultivars with improved traits such as disease resistance, yield, and quality. This review highlights the principles, applications, and advancements in embryo rescue technology in cucurbits, emphasizing its significance in cucurbit breeding programs and crop improvement efforts.

Ovarian tissue cryopreservation (OTC) and subsequent transplantation (OTT) is a fertility preservation technique widely offered to prepubertal girls and young fertile women who need to undergo oncological treatment but are at a high risk of infertility. However, OTT is not considered safe in patients with certain diseases like leukemia, Burkitt\'s lymphoma, and ovarian cancer because of the associated risk of malignant cell reintroduction. In vitro follicle development has therefore emerged as a promising means of obtaining mature metaphase II (MII) oocytes from the primordial follicle (PMF) pool contained within cryopreserved ovarian tissue, without the need for transplantation. Despite its significant potential, this novel approach remains highly challenging, as it requires replication of the intricate process of intraovarian folliculogenesis. Recent advances in multi-step in vitro culture (IVC) systems, tailored to the specific needs of each follicle stage, have demonstrated the feasibility of generating mature oocytes (MII) from early-stage human follicles. While significant progress has been made, there is still room for improvement in terms of efficiency and productivity, and a long way to go before this IVC approach can be implemented in a clinical setting. This comprehensive review outlines the most significant improvements in recent years, current limitations, and future optimization strategies.

Throughout history, parasites and parasitic diseases have been humankind\'s constant companions, as evidenced by the findings of tapeworm eggs in ancient, mummified remains. Helminths are responsible for causing severe, long-term, and debilitating infectious diseases worldwide, especially affecting economically challenged nations due to prevailing deficits in access to sanitation, proper hygiene practices, and healthcare infrastructure. Socio-ecological drivers, such as poverty, migration, and climate change, continue to contribute to parasites and their disease vectors being spread beyond known endemic zones. The study of parasitic diseases has had a fair amount of success leading to the development of new chemotherapeutic agents and the implementation of parasite eradication programs. However, further progress in this direction has been hampered by the challenges of culturing some of these parasites in in vitro systems for efficient availability, basic life cycle, infection studies, and effectiveness of novel treatment strategies. The complexity of the existing models varies widely, depending on the parasite and its life cycle, ranging from basic culture methods to advanced 3D systems. This review aims to highlight the research conducted so far in culturing and maintaining parasites in an in vitro setting, thereby contributing to a better understanding of pathogenicity and generating new insights into their lifecycles in the hopes of leading to effective treatments and prevention strategies. This work is the first comprehensive outline of existing in vitro models for highly transmissible helminth diseases causing severe morbidity and mortality in humans globally.

Walnuts (Juglans sp.) are allogamous species. Seed-derived plants are not always superior to the selected parent. Clonal propagation of selected stock plants is an essential requirement for the clonal fidelity of the descendants and to maintain their genetic structure. Selection of the desired plant is realized only after reaching maturity, and characterizing and evaluating the performance of adult trees require a long time. Clonal propagation methods ensure proper transmission of characters to descendants and can be used effectively in breeding programs. The commercialization of a cultivar or rootstock depends on the success of vegetative propagation. Walnuts, like other tree species, are recalcitrant to conventional vegetative propagation methods and even non-conventional in vitro culture (micropropagation). Elucidation of factors determining the success of cloning of desired plants would contribute to understanding current limitations for most genotypes of Juglans. We outline the role of grafting and cuttings and stool layering, as well as in vitro culture on walnut multiplication. These techniques are, in practice, entirely different; nevertheless, they are affected by common factors. The incompatibility of stock-scion and the reduced ability of stem cuttings to root are the main bottlenecks for grafting and cutting, respectively. Genotype, age, and physiological status, reinvigoration or rejuvenation-treatment of donor plant, period of harvesting and processing of explants critically affect the results of methods followed. The in vitro culture technology is the most suitable for walnut cloning. This also has constraints that affect commercial propagation of most desired genotypes. We describe comprehensive results and synthesis in this review on the asexual reproduction of walnuts, providing a better comprehension of the limiting factors and the ways to overcome them, with direct implications on commercial propagation and the releasing of outstanding genotypes.

Cacti are one of the most significant and diversified groups of angiosperms, distributed and cultivated globally, mostly in semi-arid, arid, and the Mediterranean climate regions. Conventionally, they are propagated by seeds or through vegetative propagation via rooted offshoots or grafting. However, these multiplication procedures remain insufficient for mass propagation. In vitro culture techniques are utilized to mass propagate endangered and commercial cacti species. These include somatic embryogenesis and plant regeneration through indirect or direct organogenesis. The latter is a promising tool for commercial clonal propagation of high-value species and has been successfully implemented for several species, such as Mammillaria, Hylocereus, Cereus, Echinocereus, and Ariocarpus. However, its success depends on explant type, basal nutrient formulation of culture medium, and types and concentrations of plant growth regulators. This study aimed to assess the potential of in vitro propagation methods applied to cacti species and discuss the different factors affecting the success of these methods. This study has also highlighted the insufficient work on Opuntia species for mass propagation through axillary buds\' proliferation. The development of an efficient micropropagation protocol is thus needed to meet the supply of increasing demand of Opuntia species for human consumption as fruit, animal feed, and ecological restoration in semi-arid and arid zones.

Melatonin is a natural indolamine that regulates many physiological functions in plants. The most prominent role of melatonin in plants has been its ability to work as an anti-stressor agent. Exogenous melatonin can prevent cell death and promote cell proliferation through its antioxidant properties, enhancement of polyamine biosynthesis, and the ability to shift cell metabolism in case of stressors like sugar starvation. Melatonin scavenges reactive oxygen species and thus preventing damage to cell membranes and other organelles. Its application in different plant culture systems reveals its important physiological and biochemical roles during the growth and development of these cultures. It has been observed that the exogenous melatonin protects callus culture, reduces cold-induced apoptosis in cell suspension, and stimulates adventitious and lateral roots formation. This review presents the physiological and biochemical effects of exogenous melatonin on in vitro culture systems, including its impact on biomass accumulation, growth, and development of plants.

Considerable challenges in engineering the female reproductive tissue are the follicle\'s unique architecture, the need to recapitulate the extracellular matrix, and tissue vascularization. Over the years, various strategies have been developed for preserving fertility in women diagnosed with cancer, such as embryo, oocyte, or ovarian tissue cryopreservation. While autotransplantation of cryopreserved ovarian tissue is a viable choice to restore fertility in prepubertal girls and women who need to begin chemo- or radiotherapy soon after the cancer diagnosis, it is not suitable for all patients due to the risk of having malignant cells present in the ovarian fragments in some types of cancer. Advances in tissue engineering such as 3D printing and ovary-on-a-chip technologies have the potential to be a translational strategy for precisely recapitulating normal tissue in terms of physical structure, vascularization, and molecular and cellular spatial distribution. This review first introduces the ovarian tissue structure, describes suitable properties of biomaterials for ovarian tissue engineering, and highlights recent advances in tissue engineering for developing an artificial ovary. STATEMENT OF SIGNIFICANCE: The increase of survival rates in young cancer patients has been accompanied by a rise in infertility/sterility in cancer survivors caused by the gonadotoxic effect of some chemotherapy regimens or radiotherapy. Such side-effect has a negative impact on these patients\' quality of life as one of their main concerns is generating biologically related children. To aid female cancer patients, several research groups have been resorting to tissue engineering strategies to develop an artificial ovary. In this review, we discuss the numerous biomaterials cited in the literature that have been tested to encapsulate and in vitro culture or transplant isolated preantral follicles from human and different animal models. We also summarize the recent advances in tissue engineering that can potentially be optimal strategies for developing an artificial ovary.

Cultured muscle tissue-based protein products, also known as cultured meat, are produced through in vitro myogenesis involving muscle stem cell culture and differentiation, and mature muscle cell processing for flavor and texture. This review focuses on the in vitro myogenesis for cultured meat production. The muscle stem cell-based in vitro muscle tissue production consists of a sequential process: (1) muscle sampling for stem cell collection, (2) muscle tissue dissociation and muscle stem cell isolation, (3) primary cell culture, (4) upscaled cell culture, (5) muscle differentiation and maturation, and (6) muscle tissue harvest. Although muscle stem cell research is a well-established field, the majority of these steps remain to be underoptimized to enable the in vitro creation of edible muscle-derived meat products. The profound understanding of the process would help not only cultured meat production but also business sectors that have been seeking new biomaterials for the food industry. In this review, we discuss comprehensively and in detail each step of cutting-edge methods for cultured meat production. This would be meaningful for both academia and industry to prepare for the new era of cellular agriculture.

Centaurium erythraea (centaury) is a traditionally used medicinal plant, with a spectrum of secondary metabolites with confirmed healing properties. Centaury is an emerging model in plant developmental biology due to its vigorous regenerative potential and great developmental plasticity when cultured in vitro. Hereby, we review nearly two decades of research on somatic embryogenesis (SE) in centaury. During SE, somatic cells are induced by suitable culture conditions to express their totipotency, acquire embryogenic characteristics, and eventually give rise to somatic embryos. When SE is initiated from centaury root explants, the process occurs spontaneously (on hormone-free medium), directly (without the callusing phase), and the somatic embryos are of unicellular origin. SE from leaf explants has to be induced by plant growth regulators and is indirect (preceded by callusing). Histological observations and culture conditions are compared in these two systems. The changes in antioxidative enzymes were followed during SE from the leaf explants. Special focus is given to the role of arabinogalactan proteins during SE, which were analyzed using a variety of approaches. The newest and preliminary results, including centaury transcriptome, novel potential SE markers, and novel types of arabinogalactan proteins, are discussed as perspectives of centaury research.

During the reproductive lifespan of a female, only a limited quantity of oocytes are naturally ovulated; therefore, the mammalian ovary possesses a substantial population of preantral follicles available to be handled and explored in vitro. Hence, the manipulation of preantral follicles enclosed in ovarian tissue aims to recover a considerable population of oocytes of high-value animals for potential application in profitable assisted reproductive technologies (ARTs). For this purpose, the technique of preantral follicle in vitro culture (IVC) has been the most common research tool, achieving extraordinary results with offspring production in the mouse model. Although promising outcomes have been generated in livestock animals after IVC of preantral follicles, the quantity and quality of embryo production with those oocytes are still poor. In recent years, the mare has become an additional model for IVC studies due to remarkable similarities with women and livestock animals regarding in vivo and in vitro ovarian folliculogenesis. For a successful IVC system, several factors should be carefully considered to provide an optimum culture environment able to support the viability and growth of preantral follicles enclosed in ovarian tissue. The cryopreservation of the ovarian tissue is another important in vitro manipulation technique that has been used to preserve the reproductive potential in humans and, in the future, may be used in highly valuable domestic animals or endangered species. Several improvements in cryopreservation protocols are necessary to support the utilization of ovarian tissue of different species in follow-up ARTs (e.g., ovarian fragment transplantation). This review aims to provide an update on the most current advances regarding supportive in vitro techniques used in equids to evaluate and manipulate preantral follicles and ovarian tissue, as well as methodological approaches used during IVC and cryopreservation techniques.