Beruflich Dokumente
Kultur Dokumente
Tema 2
1. Introducción
4. Clonación
1. Introducción
Cromosomas en la meiosis y en la fecundación
Figure 3.2. Chromosomes at meiosis and fertilization Two chromosome pairs of a hypothetical organism are illustrated. (Cooper y Hausman, 2004).
Primeros estadíos del desarrollo embrionario humano
El ciclo menstrual
Figure 21-17. Migration of mammalian primordial germ cells (PGCs). Alberts et al., 2008.
Estadíos de la oogénesis
Figure 21-25. The stages of oogenesis. Oogonia develop from primordial germ cells that migrate into the developing gonad early in embryogenesis. After a number of
mitotic divisions, oogonia begin meiotic division I, after which they are called primary oocytes. In mammals, primary oocytes are formed very early (between 3 and 8
months of gestation in the human embryo) and remain arrested in prophase of meiotic division I until the female becomes sexually mature. At this point, a small number
periodically mature under the influence of hormones, completing meiotic division I to become secondary oocytes, which eventually undergo meiotic division II to
become mature eggs (ova). The stage at which the egg or oocyte is released from the ovary and is fertilized varies from species to species. In most vertebrates, oocyte
maturation is arrested at metaphase of meiosis II and the secondary oocyte completes meiosis II only after fertilization. All of the polar bodies eventually degenerate. In
most animals, the developing oocyte is surrounded by specialized accessory cells that help to isolate and nourish it (not shown). (Alberts et al., 2008).
Oogénesis
Figure 19.29. The ovarian follicle of mammals. (A) Maturation of the ovarian follicle. When mature, it is often called a Graafian
follicle. (B) Scanning electron micrograph of a mature follicle in the rat. The oocyte (center) is surrounded by the smaller granulosa
cells that will make up the cumulus. (Gilbert, 2003).
Ovulación
Figure 19.31. Ovulation in the rabbit. The ovary of a living, anesthetized rabbit was exposed and observed.
When the follicle started to ovulate, the ovary was removed, fixed, and stained. (Gilbert, 2003).
2. Estadíos del desarrollo y características de
los espermatozoides
Estadíos de la espermatogénesis
Figure 21-
21-30. The stages of spermatogenesis.
spermatogenesis. Spermatogonia develop from
primordial germ cells that migrate into the testis early in embryogenesis.
embryogenesis. When the
animal becomes sexually mature,
mature, the spermatogonia begin to proliferate rapidly,
rapidly,
generating some progeny that retain the capacity to continue dividing indefinitely
(as stem-
stem-cell spermatogonia)
spermatogonia) and other progeny (maturing spermatogonia)
spermatogonia) that
will,
will, after a limited number of further mitotic division cycles,
cycles, embark on meiosis
to become primary spermatocytes.
spermatocytes. The primary spermatocytes continue through
meiotic division I to become secondary spermatocytes.
spermatocytes. After they complete
meiotic division II, the secondary spermatocytes produce haploid spermatids,
spermatids,
which differentiate into mature sperm (spermatozoa).
spermatozoa). Spermatogenesis differs
from oogenesis in several ways.
ways. (1) New cells enter meiosis continually from the
time of puberty.
puberty. (2) Each cell that begins meiosis gives rise to four mature gametes
rather than one.
one. (3) Mature sperm form by an elaborate process of cell
differentiation that begins after meiosis is complete. (4) About twice as many cell
divisions occur in the production of a sperm as in the production of an egg;
egg; in a
mouse,
mouse, for example,
example, it is estimated that on average about 56 divisions occur from
zygote to mature sperm,
sperm, and about 27 divisions occur from zygote to mature egg.
egg.
(Alberts et al., 2008).
Estructura de los tubos seminíferos de un mamífero
Figure 21-
21-29. Highly simplified drawing of a cross section of a seminiferous tubule in a mammalian testis. testis. (A) All of the stages of spermatogenesis shown take
place while the developing gametes are in intimate association with Sertoli cells.
cells. These large cells extend from the basal lamina to the lumen of the seminiferous tubule;
tubule;
they are required for the survival of the germ cells and are analogous to follicle cells in the ovary (see Figure 20-
20-18).
18). Spermatogenesis also depends on testosterone
secreted by Leydig cells,
cells, located between the seminiferous tubules.
tubules. (B) Some of these cells are self-
self-renewing stem-
stem-cell spermatogonia,
spermatogonia, whereas others are maturing
spermatogonia;
spermatogonia; after a number of mitotic divisions,
divisions, the maturing spermatogonia stop dividing by mitosis and enter meiosis to become primary spermatocytes.
spermatocytes. Eventually,
Eventually,
sperm are released into the lumen. In man,
man, it takes about 24 days for a spermatocyte to complete meiosis to become a spermatid and another 5 weeks for a spermatid to
develop into a sperm.
sperm. Sperm undergo further maturation and become motile in the epididymis;
epididymis; only then are they fully mature sperm.
sperm. (Alberts
(Alberts et al., 2008).
El espermatozoide humano
Figure 21-28. Drawing of the midpiece of a mammalian sperm as seen in cross section in an
electron microscope. The core of the flagellum is composed of an axoneme surrounded by nine
dense fibers. The axoneme consists of two singlet microtubules surrounded by nine microtubule
Figure 21-27. A human sperm. It is shown in doublets. The mitochondrion (shown in green) is well placed for providing the ATP required for
flagellar movement; its unusual spiral structure (see Figure 20-25) results from the fusion of
longitudinal section. (Alberts et al., 2008).
individual mitochondria during spermatid differentiation. (Alberts et al., 2008).
El aparato motor del espermatozoide
Figure 7.3. The motile apparatus of the sperm. (A) Cross section of the flagellum of a
mammalian spermatozoon, showing the central axoneme and the external fibers. (B)
Interpretive diagram of the axoneme, showing the "9 + 2" arrangement of the
microtubules and other flagellar components. The schematic diagram shows the
association of tubulin protofilaments into a microtubule doublet. The first ("A") portion
of the doublet is a normal microtubule comprising 13 protofilaments. The second ("B")
portion of the doublet contains only 11 (occasionally 10) protofilaments. The dynein
arms contain the ATPases that provide the energy for flagellar movement. (C) A three-
dimensional model of an "A" microtubule. The a- and b-tubulin subunits are similar but
not identical, and the microtubule can change size by polymerizing or depolymerizing
tubulin subunits at either end. (Gilbert, 2003).
El acrosoma: un tipo de lisosoma especializado
Vesí
Vesícula
Cabeza
acrosó
acrosómica
Núcleo
Porción media
Mitocondria
Membrana plasmática
Cola
Flagelo
10 μm
x 3.000
Figure 7.7. Hamster eggs immediately before fertilization. (A) The hamster egg, or ovum, is encased in the zona
pellucida. This, in turn, is surrounded by the cells of the cumulus. A polar body cell, produced during meiosis, is also
visible within the zona pellucida. (B) At lower magnification, a mouse oocyte is shown surrounded by the cumulus.
Colloidal carbon particles (India ink) are excluded by the hyaluronidate matrix. (Gilbert, 2003).
La zona pelúcida
Figure 21-22. The zona pellucida. (A) Scanning electron micrograph of a hamster egg, showing the zona pellucida. (B) A scanning
electron micrograph of a similar egg in which the zona (to which many sperm are attached) has been peeled back to reveal the underlying
plasma membrane, which contains numerous microvilli. The zona is made entirely by the developing oocyte. (Alberts et al., 2008).
Fecundación
Figure 7.20. Entry of sperm into golden hamster egg. (A) Scanning electron micrograph of
sperm fusing with egg. The "bald" spot (without microvilli) is where the polar body has
budded off. (B) Close-up of sperm-zona binding. (C) Transmission electron micrograph
showing the sperm head passing through the zona. (D) Transmission electron micrograph of
the hamster sperm fusing parallel to the egg plasma membrane. (E) Diagram of the fusion
of the sperm acrosome and plasma membranes with the egg microvilli. (Gilbert, 2003).
Desarrollo embrionario humano
Desarrollo embrionario humano
Mecanismos implicados en el reconocimiento y
fusión de los gametos. Activación del
metabolismo del cigoto
Estructura de la zona pelúcida
Expresión de ZP3 en el oocito de ratón
Figure 19.25. Expression of the ZP3 gene in the developing mouse oocyte. (A) Northern blot of ZP3 mRNA accumulation in embryonic
mouse tissues. A radioactive probe to the ZP3 message found it expressed only in the ovary, and specifically in the oocytes. (B-C) When
the luciferinase reporter gene is placed onto the ZP3 promoter and inserted into the mouse genome, the luciferinase message is seen only
in the developing oocytes of the ovary. C is a higher magnification of a section of B, showing two of the ovarian follicles containing
maturing oocytes. (Gilbert, 2003).
La reacción
acrosómica
Figure 15-40. Fertilization of an egg by a sperm triggering an increase in cytosolic Ca2+. This starfish egg was injected with a
Ca2+-sensitive fluorescent dye before it was fertilized. A wave of cytosolic Ca2+(red), released from the endoplasmic reticulum, is seen
to sweep across the egg from the site of sperm entry (arrow). This Ca2+ wave provokes a change in the egg cell surface, preventing the
entry of other sperm, and it also initiates embryonic development (discussed in Chapter 20). (Alberts et al., 2008).
La reacción
cortical del oocito
Bloqueo secundario a la
poliespermia
Figure 21-35. The coming together of the sperm and egg pronuclei after mammalian fertilization. The pronuclei migrate toward the
center of the egg. When they come together, their nuclear envelopes interdigitate. The centrosome replicates, the nuclear envelopes break
down, and the chromosomes of both gametes are eventually integrated into a single mitotic spindle, which mediates the first cleavage
division of the zygote. (Alberts et al., 2008).
Fertilización y terminación de la meiosis
Figure 14.42. Fertilization and completion of meiosis (A) Fertilization induces the transition from metaphase II to anaphase II, leading to completion of oocyte
meiosis and emission of a second polar body (which usually degenerates). The sperm nucleus decondenses, so the fertilized egg (zygote) contains two haploid nuclei
(male and female pronuclei). In mammals, the pronuclei replicate DNA as they migrate toward each other. They then initiate mitosis, with male and female
chromosomes aligning on a common spindle. Completion of mitosis and cytokinesis thus gives rise to a two-cell embryo, with each cell containing a diploid genome.
(Cooper y Hausman, 2008).
Fertilización
humana
Figure 19.21. Changes in the number of germ cells in the human ovary over the life span. (Gilbert, 2003).
Conceptos
Esterilidad
Parejas que, siendo la mujer mayor de 30 años, no han
conseguido un embarazo tras un año de relaciones
sexuales sin contracepción
Infertilidad
Parejas que consiguen una concepción pero el embarazo
se malogra y se produce una pérdida fetal
Figure 21.3. The fates of 20 hypothetical human eggs in the United States and western Europe. Under normal conditions,
only 6.2 eggs of the original 20 would be expected to develop successfully to term. (After Volpe 1987.) (Gilbert, 2003).
Edad de la madre y tasa de abortos
< 30 5%
30 - 34 7%
35 - 39 15 %
40 - 41 25 %
42 - 43 35 %
44 - 46 50 %
Miscarriage rates for women with a history of infertility tend to be higher than for fertile women.
Most (if not all) of the increased risk for miscarriage in "older" women is due to the increase in
chromosomal abnormalities (karyotype) in their eggs. This is also part of the reason for the
decline in overall fertility with aging.
Riesgo de anomalías genéticas en el recién nacido en
función de la edad de la madre
25 1 / 476
30 1 / 385
35 1 / 164
40 1 / 51
45 1 / 15
Recién nacidos vivos en función del número de oocitos
extraídos y edad de la madre
Morfología del útero y trompas de Falopio
Histerosalpingografía
Normal hysterosalpingogram
A smooth triangular uterine cavity and spill from the ends of both tubes
Morfología del útero, trompas de Falopio y ovarios
Laparoscopia
A normal left tube at laparoscopy for infertility. Course of tube is marked by "T"s
Normal fimbriated end of tube is shown at "F"
Left ovary is the white structure between the uterus and tube
El ciclo menstrual
Requisitos
Función ovárica activa
Madurez ovocitaria
Recuperación espermática de una cantidad mínima de
espermatozoides con buena movilidad (100.000/ml)
Indicaciones principales
Factor masculino moderado
Aglutinación espermática grave
Factor tubárico
Infertilidad causada por alteración de las
trompas de Falopio
Ultrasound image of an ovary at the beginning of a menstrual cycle. No medications are being given.
The ovary is outlined in blue. There are 9 antral follicles visible - marked with red spots
The ovary has normal volume (cursors measuring ovary = 30 by 17.8mm)
This woman had regular periods and a normal response to injectable FSH
Infertilidad: Ovario poliquístico
High ovarian volume and high antral follicle counts
Ultrasound image of an ovary at the beginning of a menstrual cycle. No medications are being given.
The ovary is outlined in blue. There are numerous antral follicles visible - marked with red spots
16 are seen in this image, this ovary had a total of 35 antrals (only 1 plane is shown above)
This is a polycystic ovary, with a higher than average antral count and volume (ovary = 37 by 19.5mm)
This woman had very irregular periods and was a "high responder" to injectable FSH medication
Infertilidad: Ovario poliquístico
2. Extracción de ovocitos
3. Inseminación de los ovocitos obtenidos
Inseminación clásica
Inyección intracitoplasmática de espermatozoides (ICSI)
4. Cultivo in vitro hasta embrión en diferentes estadíos de desarrollo
Cultivo durante tres días en el laboratorio
Co-cultivo embrionario durante 6 días hasta el estadío de blastocisto
- Células de la trompa de Falopio
- Células del endometrio
5. Transferencia embrionaria
En la cavidad uterina por vía transcervical
En las trompas de Falopio
6. Congelación y descongelación de embriones en su caso
Obtención de los oocitos para FIV
Ultrasound of multiple follicles (3 black circles) in a The egg aspiration procedure in progress - an egg is being
stimulated ovary aspirated from a follicle
The needle is the bright white structure (right side) above the
3rd white dot from bottom
Ovary outlined in blue, top of vagina in red
Fases de la fecundación in vitro (FIV)
- Biopsia testicular
Inyección intracitoplasmática de espermatozoides
(ICSI)
Inyección intracitoplásmica de espermatozoide (ICSI)
Requisitos
Función ovárica activa
Madurez oocitaria
Existencia de, al menos, tantos espermatozoides
móviles como oocitos se hayan obtenido
Indicaciones principales
Factor masculino severo
Alteración de membrana ovocitaria
Fallo de fecundación en FIV convencional
ICSI
High quality 8-cell embryo from in vitro fertilization Another high quality 8-cell embryo
4 cells are seen in the plane of focus 6 cells are seen in the plane of focus
We are hatching this embryo just prior to the embryo Another cell is in the center, above the plane of focus -
transfer procedure with a little imagination you can see it
The holding pipette is on the far left
Día 4: Embrión humano en estadío de mórula
A morula contains about 10 - 30 cells or so. The morula stage is the final stage prior to formation of a fluid filled cavity called
the blastocoel cavity. Once the cavitation is recognizable we call the embryo an early blastocyst.
Embryo arrest at the morula stage is not uncommon, which is one reason that transfer at the blastocyst stage can be a beneficial
IVF treatment strategy.
Día 5: Embrión humano en estadío de blastocisto
Day 5 embryo
High quality human blastocyst
Técnicas de co-cultivo embrionario
FIV con co-cultivo embrionario (1)
5. Transferencia embrionaria
En la cavidad uterina por vía transcervical
En las trompas de Falopio
6. Congelación y descongelación de embriones en su caso
Transferencia embrionaria
During the embryo transfer procedure the Wallace catheter is seen in the cervix and uterine lining (look below the yellow lines).
It is seen coming from the vagina into the cervix on the right. The catheter is also seen inside the uterine lining (to left side). The
embryos are released from the catheter tip - which is seen just to the right of the "L". Notice that the angle between the cervix
and uterus (green lines) is not severe. Cervix = C, Bladder = B, Lining = L, Vagina = V
La línea endometrial
c. Menopausia
Receptoras de oocitos (2)
B. Grupo 2
a. Anormalidades genéticas; enfermedades transmisibles a la descendencia. Se debe realizar un
consejo preconcepcional para dilucidar las posibilidades de transmisión a la descendencia:
- Autosómicas dominantes: alopecia familiar, epidermiolisis bullosa, etc.
- Autosómicas recesivas: que comparte el varón y no acepta el uso de semen de donante:
fibrosis quística, etc.
- Enfermedades ligadas al sexo: hemofilia, etc.
b. Anormalidades cromosómicas
- Mosaicismos
- Translocaciones
- Mujeres portadoras del síndrome del X- frágil
- Inversiones cromosómicas
- Deleciones, etc.
B. Grupo 2
d. Abortos de repetición
- Mala calidad ovocitaria
- Alteración cromosómica en la mujer
- Alteración cromosómica en los embriones
• Edad: 18 a 35 años
• Buen nivel intelectual
• Historial familiar negativo para enfermedades de transmisión genética
• Cariotipo normal (estudio cromosómico)
• Estudio negativo para enfermedades de transmisión sexual
SIDA
Hepatitis B y C
Clamydia
Herpes virus
Citomegalovirus
Toxoplasma gondii
Rubeola
Sífilis
• Normalidad del aparato reproductor
• Salud física y mental
• Historia de fertilidad previa y/o adecuada respuesta al tratamiento de estimulación ovárica
Recién nacidos vivos en función de la edad de la mujer
receptora de los embriones
It shows the rate of live births per embryo transfer procedure by the age of the recipient of the embryos. The blue line shows data using the infertile
woman's own eggs for IVF, while the black line shows IVF data from using donor eggs.
This chart is very useful in illustrating the decline in live birth rates by female age beginning at about age 31. This curve becomes steeper (egg quantity and
quality decreasing at a faster rate) starting at about age 37. It is very important to remember that each data point on the curve below represents an average
live birth rate from many cases. Every couple is unique and could be more fertile, or less fertile as compared to the average for their age.
Another interesting point that is illustrated here is that there is no decline in live birth rates by age of recipient when donor egg IVF is being utilized. In
other words, the age of the eggs is very important, but the age of the uterus is not.
El factor masculino:
Laboratorio de Andrología
Laboratorio de Andrología
o Concentración de espermatozoides
o Movilidad
La concentración y movilidad de los espermatozoides en las muestras son
valoradas con la ayuda de un microscopio de contraste de fases a 20
aumentos, en una cámara especial en la que no se afecta el comportamiento
de los espermatozoides
o Morfología
Tras realizar una tinción histológica adecuada, la muestra es analizada a
100 aumentos, en busca de defectos de los espermatozoides en la cabeza,
la pieza intermedia y la cola
o Viabilidad
Porcentaje de espermatozoides viables en el eyaculado
Estudio de los espermatozoides
o Ensayos inmunológicos
Detección de la presencia de anticuerpos antiespermatozoide en semen y
suero de la pareja, de los que se ha demostrado una relación directa con la
infertilidad
o Ensayos bioquímicos
Marcadores de glándulas accesorias en el aparato genital masculino, cuyo
funcionamiento es fundamental en la correcta maduración de los
espermatozoides
o Capacitación diagnóstica
Mediante esta técnica se consigue obtener el mayor número de
espermatozoides móviles posible, así como el lavado del plasma seminal
(perjudicial para los espermatozoides)
Eliminación del virus de la hepatitis C y del VIH
Eliminación del virus de la Hepatitis C en hombres Lavado de semen en varones seropositivos para el
portadores mediante lavado de semen virus de inmunodeficiencia humana
Recuperación Espermática Extrema
Características
Técnica empleada en el tratamiento de pacientes azoospémicos, es decir,
pacientes que carecen de espermatozoides en el eyaculado
Técnica biópsica
Extracción de una pequeña muestra de tejido testicular
Requisitos
Existencia de espermatozoides en número mínimo
Indicaciones principales
Azoospermia obstructiva
Azoospermia no obstructiva o secretora
Imposibilidad de recoger la muestra
Inmovilidad absoluta de espermatozoides en el eyaculado
Banco de semen
Las muestras de semen se conservan congeladas, usando como refrigerante
nitrógeno líquido, técnica que se conoce como criopreservación
Pacientes
- Congelaciones pre-vasectomía
- Pacientes sometidas a tratamientos de reproducción asistida cuyas
parejas, por motivos laborales, no puedan estar presentes en el
momento en el que se necesita su semen
- Congelaciones previas a tratamientos de quimio- o radioterapia
- Dificultades en la obtención del eyaculado
- Muestras de muy mala calidad
- Biopsias testiculares
- Aspiraciones de epidídimo
Banco de semen
Donantes
- Antígenos de la Hepatitis B
- Anticuerpos anti-HIV
- Chlamydia
- Anticuerpos anti-herpes virus
- Sífilis
- Anticuerpos anti-Hepatitis C
- Gonorrea
- Citomegalovirus
Utilización de semen de donante
Técnica
Marcaje con fluorescencia de los cromosomas con sondas de ADN específicas
para los cromosomas motivo de estudio
Aplicaciones
Hibridación in situ fluorescente (FISH). Consiste en marcar con fluorescencia los cromosomas con sondas de ADN (ácido
desoxiribonucleico) específicas para los cromosomas motivo de estudio. A continuación, con el microscopio de fluorescencia,
podemos identificar el número de copias para un cromosoma determinado.
FISH
Técnica
Consiste en la amplificación de secuencias específicas del ADN de un gen,
en las que la presencia de una mutación desencadena una enfermedad de
origen génico
Aplicaciones
Se han descrito aproximadamente 5.000 enfermedades de origen génico,
como son:
Fibrosis quística
Distrofia miotónica
Enfermedad de Tay-Sachs
Síndrome de Marfan
Beta-talasemia
Anemia falciforme
Enfermedad de Huntington
Etc.
Reacción en cadena de la polimerasa (PCR). Consiste en la amplificación de secuencias específicas del ADN de un gen, en las que la
presencia de una mutación desencadena una enfermedad de origen génico. Nos permite diferenciar qué embriones son normales y cuáles
tienen un gen mutado y, por tanto, desarrollarán la enfermedad.
Enfermedades humanas detectables mediante
diagnóstico pre-implantacional
Autosómicas recesivas
Fibrosis Quística (gen CFTR)
Atrofia Muscular Espinal (Tipo I o de Werdnig-Hoffmann) (gen SMN1, Deleción exon 7)
Talasemia (25-26delAA, IVS2+1G>A, IVS1+6T>C, IVS1+110G>A)
Anemia Falciforme
Incompatibilidad factor Rhesus D. (Duplex RhD y RhCE)
Autosómicas dominantes
Distrofia Miotónica o Enfermedad de Steinert
Enfermedad de Huntington
Enfermedad Charcot-Marie-Tooth (gen MPZ)
Enfermedad AD Riñón poliquístico
Síndrome de Marfan (MFS) (gen fibrillin-1 [FBN1])
Ligadas al cromosoma X
Síndrome del X frágil
Distrofia Miotónica de Duchenne (DMD) y DM Becker (DMB)
Hemofilia A
Detección de anomalías cromosómicas
An egg with a severely fragmented polar body An egg with many pronuclei (hard to see here)
at 10 to 1 o'clock and severe fragmentation
This is quite unusual This is rare and consistent with a chromosomal
abnormality
Oocitos anormales
3 degenerative eggs Close detail of one of the degenerative eggs from the same
This patient also had some "normal" eggs case as shown above
Óvulo humano fertilizado anormal
Presencia de tres pronúcleos
Normal Abnormal
Chromosomes line up in Chromosomes line up
straight line on spindle erratically
D.E. Battaglia, et al. (1996) Influence of maternal age on meiotic spindle assembly in
oocytes from naturally cycling women. Human Reproduction, Vol. 11:2217-2222.
Oocitos y embriones humanos con alteraciones en
la zona pelúcida
These low quality blastocysts are shown to indicate that blastocyst culture is not a magic bullet for infertility. Most human embryos
do not have the genetic potential to hatch from their shells, implant and continue normal development in order to result in a live birth.
4. Clonación
Clonación
Transferencia nuclear
somática
Freddy
Figure 4.5. Procedure for transplanting blastula nuclei into activated
enucleated Rana pipiens eggs. The relative dimensions of the meiotic
spindle have been exaggerated to show the technique. "Freddy," the
handsome and mature R. pipiens in the photograph, was derived in this
way by M. DiBerardino and N. Hoffner Orr. (Gilbert, 2003).
Transferencia nuclear somática
Figure 4.6. Percentage of successful nuclear transplants as a function of the developmental age of the donor nucleus.
The abscissa represents the developmental stage at which a donor nucleus (from R. pipiens) was isolated and inserted
into an activated enucleated oocyte. The ordinate shows the percentage of those transplants capable of producing
blastulae that could then direct development to the swimming tadpole stage. (Gilbert, 2003).
Clonación
Clonación a partir de núcleos de
células diferenciadas
Figure 4.7. A clone of Xenopus laevis frogs. The nuclei for all
the members of this clone came from a single individual a
female tailbud-stage tadpole whose parents (upper panel) were
both marked by albino genes. The nuclei (containing these
defective pigmentation genes) were transferred into activated
enucleated eggs from a wild-type female (upper panel). The
resulting frogs were all female and albino (lower panel).
(Gilbert, 2003).
Clonación de
mamíferos
Figure 4.18. Insertion of new DNA into embryonic cells. Here, DNA (from cloned genes) is injected into the a pronucleus
of a mouse egg. (Gilbert, 2003).
Obtención de animales transgénicos
Figure 4.10. Production of transgenic sheep. The structural gene for an important human protein (such as clotting factors, insulin, or
a1-antitrypsin) is linked to the regulatory region (promoter) of a sheep milk protein gene (such as that for casein or lactalbumin). This
recombinant gene is injected into the pronucleus of a newly fertilized sheep egg, and the egg is implanted into a foster mother. The
newborn sheep are screened for the presence of the human gene. When the transgenic sheep mature, the human gene should be
expressed in the mammary gland and the protein secreted into the milk. From the milk one can then isolate large amounts of the
protein for pharmaceutical use. (Gilbert, 2003).
Prometea
El primer caballo clonado del mundo
C. Galli, 28-5-2003
Science, 2000
Oregon Primate
Research Center
Objetivo: crear nuevos modelos animales para el estudio y tratamiento de enfermedades humanas
Noel, Angel, Star, Joy, Mary
Primeros cerditos creados para realización de xenotransplantes
Alan Colman, PPL Therapeutics, Virginia, 25-12-2001
Células madre y clonado terapéutico
Preparación de células madre embrionarias
Clonación reprodutiva y terapéutica