Organoides: fundamentos, presente y futuro / Organoids: fundamentals, present and future

RESUMEN Los organoides son estructuras miniaturizadas, generadas principalmente a partir de células madre pluripotentes inducidas, que se cultivan en el laboratorio conservando sus características innatas o adquiridas. Tienen el potencial de reproducir procesos de desarrollo biológico, modelar procesos patológicos que permitirán el descubrimiento de nuevos fármacos y propicien la medicina regenerativa. Sin embargo, estas experiencias requieren perfeccionamiento constante porque pueden haberse realizado variaciones en la constitución de estos órganos. Por ello, el presente artículo tiene como objetivo revisar la información actualizada sobre organoides y sus procesos experimentales básicos y recientes, empezando por la gastrulación, para tratar de imitar, en lo posible, la formación de las tres capas: ectodermo, mesodermo y endodermo, incluyendo los factores que intervienen en la inducción, diferenciación y maduración en la generación de estos organoides. Asimismo, el diseño y preparación de medios de cultivo altamente especializados que permitan obtener el órgano seleccionado con la mayor precisión y seguridad. Se realizó una búsqueda de artículos originales y de revisión publicados en PubMed, Nature y Science. Los artículos se seleccionaron por sus resúmenes y por su texto completo. Las conclusiones de este articulo destacan las ventajas futuras en el uso y aplicaciones de los organoides.

ABSTRACT Organoids are tiny structures, mainly generated from induced pluripotent stem cells, which are cultured in the laboratory while retaining their innate or acquired characteristics. They have the potential to reproduce biological development processes, model pathological processes that will enable the discovery of new drugs and promote regenerative medicine. However, these processes require constant improvement because variations may have occurred in the constitution of the organs. Therefore, this article aims to review updated information on organoids and their basic and recent experimental processes, starting with gastrulation, in an attempt to mimic, as much as possible, the formation of the three layers: ectoderm, mesoderm and endoderm; as well as the information regarding the factors involved in the induction, differentiation and maturation during the generation of organoids. Likewise, the design and preparation of highly specialized culture media that allow obtaining the selected organ with the highest precision and safety. We searched for original and review articles published in PubMed, Nature and Science. Articles were selected for their abstracts and full text. The conclusions of this article highlight the future advantages in the use and applications of organoids.

Holoprosencefalia semilobar y malformaciones asociadas: reporte de caso y algunas consideraciones / Semilobar holoprosencephaly and associated malformations: case report and some considerations

RESUMEN: El defecto más común del prosencéfalo es la holoprosencefalia (HPE), caracterizada por ausencia en la división del prosencéfalo. La holoprosencefalia tiene una prevalencia de 1/10.000 en recién nacidos; la ciclopía de 1/100.000 nacidos y la agnatia asociada a holoprosencefalia de 0,8 a 10 %. El objetivo fue describir las características morfológicas e histopatológicas de un feto humano con holoprosencefalia y sus malformaciones asociadas. Se estudió un feto masculino. Se le realizó microdisección bajo el estereomicroscopio, toma de microfotografías con cámara AxioCam y software AxioVision 4.8, y estudio histopatológico. La edad gestacional estimada fue de 12,4-13,2 semanas, encontrándose como hallazgos la HPE semilobar asociada a ciclopía, esbozo oral hipoplásico sin apertura oral, cubierta por una membrana y ausencia de labios. El estudio histopatológico reportó: ojo con lente, retina y córnea únicos; en la cara, probóscide con cartílago tubular en formación asociado a mesénquima y cubierta muscular esquelética, y cavidad oral pequeña, circunscrita por mandíbula hipoplásica conformada por cartílago. Se revisa la literatura y se reafirma la necesidad de estudio multidisciplinario de esta patología para mejorar su comprensión.

SUMMARY: The most common defect of the forebrain is holoprosencephaly (HPE), characterized by absence in the forebrain division. Holoprosencephaly has a prevalence of 1 / 10,000 in newborns; the cyclopia of 1 / 100,000 births and the agnathia, in a series of cases of holoprosencephaly ranges from 0.8 to 10 %. The objective was the description of the morphological and histopathological characteristics of fetus with holoprosencephaly and its associated malformations. A male fetus was studied. Microdissection was performed under the stereomicroscope, taking microphotographs with AxioCam camera and AxioVision 4.8 software, and histopathological study. The estimated gestational age was 12.4-13.2 weeks, the findings were semilobar HPE, associated with cyclopia, hypoplastic oral outline without buccal opening, covered by a membrane and lips absence. The histopathological study reported: eye with lens, retina and cornea only; in the face, proboscis with tubular cartilage in formation associated with mesenchyme and musculoskeletal sheath, and small oral cavity, delimited by hypoplastic mandible conformed by cartilage. The literature is reviewed and reaffirmed the need for multidisciplinary studies of this disease to improve their understanding.

ism1 regulates convergence and extension movements during zebrafish gastrulation / 中国比较医学杂志

Objective During zebrafish embryo gastrulation, the embryonic body plan is established by coordinated movements,including epiboly,involution,and convergence and extension(CE)movements. The aim of this study was to examine the expression patterns and the developmental functions of isthmin 1(ism1)during zebrafish gastrulation. Methods First, the expression of ism1 was examined using whole mount in situ hybridization during zebrafish embryonic development. Next,antisense morpholino oligonucleotide against ism1 and ism1 mRNA were injected into zebrafish embryos,respectively,to investigate its role in CE movements. Results ism1 is an early zygotic gene that is expressed in the dorsal side at 30% epiboly stage and in the blastoderm margin with the highest level in the dorsal organizer at shield stage as revealed by whole-mount in situ hybridization experiments. Compared with the wild-type embryos,either gain-or loss-of-function of ism1 leads to severe defects of CE movements. Conclusions Ism1 plays an essential role in CE movements during zebrafish embryo gastrulation.

Understanding of Human Embryo Development for Teratogen Counselling

Human embryology is the study of development from a single cell to a baby in 9 months. Implantation occurs at the end of the first week of development. The second week of development is known as the week of 2's. Gastrulation, the most characteristic event occurring in the third week, establishes three germ layers composed of ectoderm, mesoderm, and endoderm. The three germ layers and neural crest cells lead to the development of their own tissues and organs during the embryonic period, which extends from the third to the eighth week. Major congenital malformations occur in the embryonic period. The fetal period, from the third month to the day of birth, is the time for maturation of tissues and organs, and growth of the body. Because of the close relationship between embryology and congenital abnormalities, knowledge of human development is essential to assess the effects on the embryo when the mother has been exposed to teratogens. This paper briefly reviews the normal embryonic development and associated congenital malformation.

Desarrollo embrionario del pargo colorado Lutjanus colorado (Jordan & Gilbert, 1882) / Embryonic development of pargo colorado Lutjanus colorado (Jordan & Gilbert, 1882)

El pargo colorado (Lutjanus colorado) es una especie con un alto valor comercial en el mercado mexicano, con potencial para su cultivo. Hasta la fecha no existen estudios sobre su reproducción, cultivo larvario y engorda en cautiverio. El presente trabajo es el primer reporte sobre la descripción a detalle del desarrollo embrionario de la especie bajo condiciones de cultivo. Los huevos fertilizados viables del pargo colorado son pelágicos, esféricos, transparentes y con una sola gota de aceite. Midieron 0,77±0,09 mm de diámetro y la gota de aceite 0,14±0,01 mm. La primera división ocurrió a las 0,05 horas post fertilización (HPF). La eclosión se llevó a cabo a las 17,22 HPF bajo las condiciones del presente estudio. Las larvas recién eclosionadas midieron 1,8±0,1 mm de longitud total (LT). El desarrollo embrionario de esta especie fue similar a la descrita para especies de la misma familia. Los resultados del presente estudio aportan información básica para iniciar el desarrollo de la biotecnología para la producción de semilla de esta especie a escala comercial.

The Colorado snapper (Lutjanus colorado) is one of the most commercially important fish species in México and it is considered a suitable candidate for culture. Until now, no research has been carried out on its reproduction, larviculture and fattening in captivity. This study is the first description of embryonic development of this species under controlled conditions. Fertilized eggs of Colorado snapper are pelagic, spherical and transparent and contain one drop of oil. Eggs measured 0.77±0.09 mm and the drop of oil 0.14±0.01 mm. First cell division occurred at 0.05 h post-fertilization (HPF), hatching at 17.22 HPF under the above described conditions. Larvae total length (LT) was 1.8±0.1 mm. Embryonic development of this species was similar to other lutjanidae species. These results provide basic information for developing the necessary biotechnology for commercial seed production of the Colorado snapper.

Desarrollo embrionario del capaz pimelodus grosskopfii (Steindachner, 1879) / Embryonic development of capaz pimelodus grosskopfii (Steindachner, 1879)

El conocimiento del desarrollo embrionario en los peces es especialmente importante en especies nativas con potencial para la piscicultura, en virtud que permite identificar eventos morfológicos y cronológicos, necesarios para establecer prácticas de manejo durante las fases de incubación y larvicultura. El capaz (Pimelodus grosskopfii) es una especie con potencial para cultivo comercial, por sus hábitos alimenticios omnívoros y aceptación de su carne en el mercado. Para estudiar el desarrollo embrionario de la especie, ejemplares adultos sexualmente maduros fueron inducidos a la reproducción con extracto de hipófisis de carpa (5,75 y 4,0 mg Kg-1, hembras y machos, respectivamente). Los óvulos seminados fueron incubados en un sistema de flujo ascendente de 30 L a 27 +/- 1 °C. Las muestras (n=30) fueron colectadas al momento de la extrusión, durante la fertilización y cada 15 minutos a partir de las 0 horas postfertilización (HPF) hasta las 2 horas y cada 30 minutos desde las 2 HPF hasta 5 HPF; finalmente, entre las 5 HPF y la eclosión, cada 60 minutos. Los óvulos fertilizados presentaron forma esférica, sin adherencias y con amplio espacio perivitelino. El desarrollo embrionario finalizó a las 12 HPF. La diferenciación del polo animal y vegetal ocurrió a las 0,2 HPF, el primer clivaje a las 0,3 HPF, el blastodisco alto y estratificado a las 1,8 HPF, el blastodisco achatado a las 3,3 HPF, la epibolia < a 50 por ciento se observó a las 4 HPF, el cierre del blastoporo a las 5,7 HPF, la diferenciación cráneo caudal e inicio de la neurolación a las 7 HPF, la diferenciación de las vesículas ópticas, óticas y vesícula de Kupffer a las 8,5 HPF, la liberación de la cola del vitelo a las 10 HPF, los primeros movimientos se observaron a las 10,5 HPF y finalmente la eclosión ocurrió a las 12 HPF. Las larvas al eclosionar presentaron una longitud total de 2987+/-67 um, sin pigmentación, tracto digestivo rudimentario, sin abertura bucal ni anal y presencia de cromatóforos...

The knowledge of embryonic development in fish is important in native species with potential for fish farming, by virtue of which it makes possible to identify morphological and chronological events to establish management practices during incubation periods and larviculture. The capaz (Pimelodus grosskopfii) is a species with potential for commercial crop, due to their omnivorous eating habits and acceptance of its meat in the market. To study the embryonic development of the species, sexually mature adult specimens were induced to reproduce with carp pituitary extract (5.75 and 4.0 mgKg-1, females and males, respectively). The inseminated oocytes were incubated in an upward flow system 30 a 27 +/- 1 ° C. The samples (n = 30) were collected at the same time of the extrusion, during fertilization, and every 15 minutes starting from 0 to 2 hours post fertilization (HPF) and every 30 minutes from 0 to 2 HPF, and every 30 minutes from 2 to 5 HPF; finally, between 5 HPF and hatching every 60 minutes. The fertilized oocytes had a spherical shape without adhesions and large perivitelline space. Embryonic development took 12 HPF. The differentiation in animal and vegetal pole occurred at 0.2 HPF, the first cleavage at 0.3 HPF, stratified and high blastodisc at 1.8 HPF, flattened blastodisc at 3.3 HPF, the epiboly <50 percent was observed at 4 HPF, the closure of the blastopore at 5.7 HPF, cranial-caudal differentiation and starting the neurolation at 7 HPF, the differentiation of the optic vesicles, otic and Kupffer's vesicle at 8.5 HPF, tail of the vitelum was released at 10 HPF, first movements were observed at 10.5 and finally hatching occurred at 12 HPF. When the larvae hatched, they showed a total length of 2987+/-67 µm, without depigmentation, rudimentary digestive system without oral and anal opening and the presence of chromatophores on the yolk sac.

Desarrollo embrionario del sistema nervioso central y órganos de los sentidos: revisión / Embryonic central nervous system and sense organ development: review

El sistema nervioso tiene origen en la capa germinal ectodérmica. Así como en la vida posnatalel sistema nervioso está claramente diferenciado en sistema nervioso central y periférico,en su etapa embrionaria la formación de cada uno sigue caminos diferentes, pero cercanos.Esta revisión hace hincapié en la formación del sistema nervioso central y de estructurasespecíficas, como los órganos de los sentidos. Se presenta un breve recorrido por procesoscomo la gastrulación, la neuralización y la formación de placodas hasta llegar a las diferentesestructuras que conforman el sistema nervioso central y los órganos de los sentidos...

The nervous system has its origin in the well-known ectodermic germinal layer. During thepostnatal life, the nervous system is clearly differentiated in central nervous system andperipheral nervous system. During the embryonic stage, all the components follow differentpaths but they are still close to each other. This review focuses on the central nervoussystem and development of the sense organs. It includes a brief review of processes suchas gastrulation, neurulation and development of cranial placodes and the structures thatmake up the central nervous system and the sense organs...

The Effects of Retinoic Acid on Gastrulation of Hatched Mouse Embroyos / 대한산부인과학회잡지

Retinoic acid(RA), formed in vivo by oxidation of retinol, is known as morphogenic signal. RA plays an active role in normal embryonic development at physiological concentration, but excess RA can be a powerful teratogen in human and animals. The present study was designed to examine the direct effect of RA on murine embryogenesis(gastrulation) and to define the specific development processes perturbed by RA. Five to fifteen blastocysts were randomly assigned to separate culture dishes of the experimental group. Various concentrations of RA(10(-9) M, 10(-7) M, and 10(-5) M) were used in culturing blastocysts. In the effect of RA on the normal grouwth of embryo, the rates of development to the stages of attachment, early egg cylinder(EEC), late egg cylinder(LEC), and early somite(ES)were significantly(p < 0.01) decreased as the RA concentration increased. Stil in the yolk sac formation rate, there was a significant, dose-dependent difference(p < 0.01) according to the RA concentration. In the degeneration of embryos by RA, the effect was more apparent as the concentration of Ra increased. The production rates of embryos devoid of egg cylinder region and embryos with abnormal egg cylinder region were increased (p < 0.01)in a dose-dependent manner according to RA concentration. In conclusion, RA probably act as teratogen at gastrula stage embryos in high concentration and effect of teratogenesis is dose-dependent.

THE GASTRULATION OF MONGOLIAN SHEEP EMBRYOS / 解剖学报

The Mongolian sheep embryo develops into gastrula from 8 days 17 hours 10 minutes to 12 days 14 hours 30 minutes after coitus. The blastocyst is spherical in shape about 266—972?m in diameter. The zona pellucida has disappeared. The trophoblast consists of simple flat epithlium or simple cuboidal epithelium. About 8 days 17 hours 10 minutes to 9 days 15 hours 18 minutes after coitus the embryonic knot becomes spheroid, about 70—72?m in diameter. The embryonic knot is slightly protruding from The surface of the blastocyst and is covered by the trophoblast, the ectodermal cells of the dorsal portion of the embryonic knot form a mass. The lower surface of the embryonic knot has differentiated into the endoderm. The extraembryonic endoderm extends to the inner surface of the trophoblast from the periphery of the embryoblast. About 10 days 16 hours to 10 days 17 hours 20 minutes after coitus, in the dorsal portion of the embryonic knot forms a vesicle which is known as the primitive amniotic cavity about 40?m in diameter. The bottom of the primitive amniotic cavity has differentiated into the ectoderm, about 72—90?m in diameter. The endoderm of the lower surface of the embryonic knot continues to develop along the inner surface of the trophoblast to the equator. The developmental process of the primitve amniotic cavity of Mongolian sheep embryos is different from rabbit、guinea pig and human embryos, but it is similar to the mole embryos. About 11 days 16 hours 5 minutes to 12 days 14 hours 30 minutes after coitus, the embryonic knot becomes discal in shape and is fully developed, about 252—396?m in diameter. The primitive amniotic cavity of the dorsal portion of the embryonic disc has disappeared. The ecitoderm is exposed to the surface of the blastocyst, it consists of stratified columnar epthelium. The endoderm of the lower surface of the embryonic disc consists of the simple flat epithelium, it forms a spherical primitive gut cavity along the inner surface of the trophoblbst.