Transcribed by Albert Cheng Craniofacial Biology Lecture 1 General Embryology I

3/12/14

Slide 1: General Embryology [Dr. Craig] Let me welcome you to your first lecture in Craniofacial Biology (CFB). I think youll like this course because most of the dental stuff thats in the basic science curriculum is in this course. The course is organized into 6 units. Where possible, each unit starts off with the developmental biology of the tissue or the system of interestgoes on to mature tissuefollows up with something called clinical case presentation (CCPs), which is done by a clinician usually to reinforce the clinical relevance of the unit and then culminates with a quiz. Ive already been contacted about the course policy and quizzes. Quizzes, we dont make up quizzes because its really difficult to make a really high-quality multiple choice exam. What we do in this course is you have 6 quizzes and we drop the lowest just in case you have a contingency that you cant get around (death in the family etc). Ive also been asked, can I take the quiz before and can I take the quiz after, I wish we could say yes but in the past weve had security problems. Most of your questions will probably be answered if you look at the course syllabus which is posted on the NYU courses site and the faculty that put together this course worked very hard on itmaking sure it was clinically relevant and I hope you should enjoy it and theyre real proud of this course. If you have any questions or concerns, please contact me at my email. Im usually fairly responsive. Good luck tomorrow. You have a midterm tomorrow. Thats why most of you are not here. Enjoy the course. [Dr. Saint-Jeannet] Alright still some technical issues butI think its good. Was attendance required in the last class? So nothing personal I guess. So Im gonna give you the first three lectures in this course which are primarily dealing with general concepts of embryology, early development of the human embryo. So nothing to do really with teeth in general unlike what Dr. Craig said. Ill be covering most of the primary, whats related to early embryoorganogenesis of the developing human embryo Slide 2: Recommended Textbook The textbook that I use to prepare those lectures is the textbook that I listed in the syllabus. Its available online. Its the Medical Embryology of Langman. So they are now on their 12th edition but any of the earlier edition is as good as the 12th. Theres very little change in the most recent edition as compared to the later one. Slide 3: General Embryology I The first hour we are going to talk about when everything starts. Development of an embryo starts by fertilization and that involves the two gametes, male and female. We are going to talk first about the two gameteswhere they come fromhow they formand how fertilization takes place and how we go from fertilization to the first stage which is blastocyst formation. We are going to talk about primordial germ cellswhat they arewhich are the precursors for the gametes (oocyte and sperm). Then, were going to talk about gametogenesishow you go from primordial germ cells to an oocyte or sperm. Talk about the process that is involved in generating those gametes which is meiosis, which is very specific for the primordial germ cells. Then, we talk about more of the characteristics related to the formation of the female versus male gametes.

Then, we talk about fertilization and the next step will be about cleavage and then the blastocyst formation and implantation. Slide 4: Timeline of Human Development So here this is a timeline of human development which can be divided into 3 trimesters. The birth usually takes place around 38th week. The development of the human embryo can be divided into two major phases: one is called embryonic period which corresponds to the step of fertilization all the way to 8th week of development and then the fetal period which will be essentially covering the rest of the period2nd and 3rd trimester. So most of the important stuff taking place in the development of the human embryo is taking place in the embryonic period which is sometimes referred to as the organogenesis period, where most of the organ systems start to be put in place. Today, for the first hour, we are gonna talk primarily whats taking place during the 1st and 2nd week. Well see that a lot is going on there. Slide 5: Primordial Germ Cells Development begins with fertilization which involve the fusion of haploid gametes, the oocyte and sperm which are shown here. The primary function of the gametes is to transmit the genetic information. Gametes are derived from cells that are call primordial germ cells. You can see immediately there is quite a substantial difference between the two gametes regarding the size. Slide 6: They both arise from a common precursor, the primordial germ cells (PGCs). Those PGCs are formed during the 2nd week of embryogenesis. They arise from the inner cell mass and the epiblast more specifically. Those PGCs arise from the inner cell mass of the epiblast and then move during the 3rd week of embryogenesis into the yolk sac, more specifically into wall of the yolk sac. Slide 7: This is another representation here showing you the embryo and those PGCs that are in the yolk sac here that will eventually during the 4th week migrate into the genital ridge, which will be the future gonad of the developing embryo. Those PGCs that are found in the posterior region of the yolk sacthis is the anterior part of embryothis is the posterior herethose PGCs in the yolk sac will migrate along the hindgut in this region and migrateHeres a different viewyou see those PGCs in red that are migrating along the hindgut through the mysentery here and migrate into the genital ridge, which are the future developing gonad of the embryo. So during the whole process, the PGCs that are found in the yolk sac initially start to migrate and as they migrate, they proliferate very extensively. Once they reach the genital ridge where they will settle, they continue to proliferate for a little bit...for a period of time. The time that which the PGCs start to populates the genital ridge is taking place around the 6th week of gestation. Slide 8: This is a higher magnification of the region of the genital ridge (refer to slide 7) where we are looking at those PGCs that have migrated along the hindgut and mysentery and started to settle into this genital region, which is a part of the intermediate mesoderm and we are going to talk about that from which those PGCs will eventually develop. The sex of the embryo is determined genetically at fertilization. However, the gonads do not acquire the male or female characteristics

only until the 7th week. Upon the arrival of the PGCs into the genital ridge, like I said they continue to proliferate for a little awhile, and then they will migrate into the underlying mesenchyme to form those structures here known as primitive sex cords. At this stage, when they migrate and populate the primitive sex cord, its impossible to differentiate whats a male versus a female gonad. For that reason, this gonad at this stage is referred to as an indifferent gonad. Slide 9: So what will make a testis versus an ovary is all about the genetic composition of the embryo. So in a genetically male embryo, which will have an XY sex chromosome complex, the Y chromosome carries a gene that is known as the SRY gene which is the testis-determining factor. Under the influence of SRY, the primitive sex cords will start to differentiate and develop in those now so call testis cords and give rise eventually to the testis. In the female, which have an XX sex chromosome complex, there is no SRY and in the absence of SRY, those primitive sex cords start to degenerate and what happens is that those PGCs move into the superficial epithelium where theyand the epithelium under the influence of PGCs start to proliferate and the surface epithelium will give rise to cortical cords which will give rise to the ovary itself Slide 10: So to summarize, when the PGCs arrive in the gonad, its impossible to discriminate male from female gonad, so this is known as the indifferent gonad. Its only under the influence of the SRY gene that the indifferent gonad will become a testis. So its often seen that the development of the ovary is the default pathway in the sense that in the absence of any other influence, in this case the SRY gene, you will form an ovary. So this is essentially the default stage of the gonad. Slide 11: 2-Meiosis So when you talk about gametes and PGCs, there is an important process that is exclusively associated with those cells, which is the process of meiosis. Meiosis has two important functions. You may have heard about meiosis already, you heard about mitosis. Meiosis is specific to germ cells. The two major functions of meiosis are the meiotic recombination and the reduction of the number of chromosomes to a haploid complement. Meiotic recombination is exchange of segments of chromosomes during the pairing of the chromosome. The key event of the meiosis is one round of pre-meiotic DNA replication followed by a unique prophase I stage of meiosis where the genetic exchange will take place. And two rounds of chromosome segregation referred to as meiosis I and meiosis II during which there is a reduction of the number of chromosomes to a haploid complement. So when the PGCs initiate meiosis, the PGCs is now referred to as gametes. Slide 12: Mitosis I have this slide here to remind you what mitosis is. Mitosis is the division of cell into two daughter cells that are genetically identical; they will have the same complement of 46 chromosomes. So there is replication of DNA, chromosomes align at the equatorial plate during metaphase and then those chromosome will separate during anaphase/telophase to give rise to 2 daughter cells that are identical.

Slide 13: Meiosis Meiosis is a little bit different. Whats taking place during meiosis, there is homologous chromosome will start to pair and condensate here, then they will form chiasm where the D stage, the exchange of different part of the chromosome will take place. Then, it goes through anaphase of 1st meiotic division where there is segregation of the two homologous chromosomes and then eventually give rise to two cells that have haploid complement of chromosomes. Then, the second division which will give rise to the four cells that are haploid complement of chromosomes also. Slide 14: Prophase I of Meiosis The important phase is the prophase I of meiosis which I have indicated where this genetic recombination is exchange between maternal and paternal alleles are taking place. This phase can be divided into 4 phases: leptotene, zygotene, pachytene, diplotene. The leptotene is the condensation of chromosomes. Zygotene is the pairing of those homologous chromosomes. Pachytene is the phase during which there is a crossover that takes place which is shown here, where theres exchange of chromosome. Diplotene, the synapsed chromosomes begins to separate. The exchange of genetic material is taking place and those chromosomes are starting to separate from one another. You will see this is an important phase during oogenesis. Most of oocytes are stuck in this diplotene phase for a quite a long period of time. Slide 15: Production of gametes in female and male So this is some of the difference between male and female regarding the production of gametes. So the first division of maturation which takes placewhich gives rise to 2, in the male, spermatocyte that are essentially identical that have haploid complement of chromosome. In the female, the first maturation division gives rise to oocyte and small cells which contain the haploid complement of chromosomes that will degenerate eventually. The second maturation division in the male, the secondary spermatocyte will be divided into four spermatids which will give rise to four gametes. In the female, the second maturation division, the oocyte will be divided into mature oocyte and give rise to second polar bodies. So one of the major differences here between male and female in meiosis is: female you have the production of one mature oocyte whereas in the male, you have formation of four mature gametes. Slide 16: Chart Some of the differences are also shown here. In the male genital ridge, the germ cells are committed to spermatogenesis. Sertoli cells, which are supporting cells that are producing a number of hormones that allow for the differentiation of the testis, are differentiated. The germ cells enter mitotic arrest and form spermatogonia. Mitosis is not resumed in the male until after birth and I will show you a diagram to illustrate that. The first meiotic division is initiated in the male at puberty. In female, the germ cells are committed to oogenesis. All germ cells at the same time enter meiotic prophase and become oocytes. Undergo the early stages of meiosis which is meiotic recombination and are arrested at this diplotene prophase I at the time of birth. The first meiotic division is only completed at puberty.

Slide 17: Timing of meiosis onset differs in male and female Those differences here that are highlighted between female versus male germ lines. Whats happening in the fetus, newborn, puberty, and the adult. The first meiotic division is initiated in fetus, in female, and is completed only at puberty. The completion of second meiotic division only takes place at the time of fertilization when the sperm enters the oocyte. In male, the first meiotic prophase division starts at puberty and the 2nd meiotic division takes place throughout the adult life. Slide 18: Oogenesis Some of the process associated with oogenesis, which is the process by which oogonia differentiate into mature oocytes. So once the PGCs arrive in the gonad of a genetic female, they differentiate into oogonia. Those oogonia when they arrive in the ovary will go through mitotic division; proliferate very extensively and those are the oogonia and thats the surface epithelium of the ovary here. Subsequently, around the 7th month of gestation, those oogonia are now called primary oocytes and they are in the prophase I of first meiotic division and they start to be fully surrounded by follicular cells that are derived from the surface epithelium of the ovary. The primary oocytes in the newborn are surrounded by follicular cells and those oocytes are arrested in this diplotene stage where genetic recombination has taken place but the first meiotic division has not been completed yet. Slide 19: Primary oocytes become individually surrounded by a layer of follicular epithelial cells. At this stage, they are referred to as primordial follicles. At puberty, whats happening is that a selected number of primary oocyte will grow and the follicular cells that are surrounding the primary oocyte will start to proliferate and form this stratified epithelium surrounding the primary oocyte. Those are referred to as primary follicles. Those follicular cells or granulosa cells as they are sometimes called are also involved in the production of a layer of glycoprotein that will coat the oocyte itself, which is shown in red here which is known as the zona pellucida. We will talk about that; it has a very important role in fertilization. So the primary follicle will become a secondary follicle when the cavity starts to form in the granulosa cells and this cavity here will accumulate, which is known as the antraum, liquid that is secreted by the granulosa cells. Slide 20: This is another secondary follicle which has follicular antrum here and the mature follicle which is called ovulatory follicle or graafian follicle sometimes. It is a very large structure that has a very large cavity here which is known as the antrum. There is a layer of follicular cells and the oocyte is still surrounded by the zona pellucida and a layer of follicular cells that is known as the cumulus oophorus. Slide 21: Typically, at birth, females have approximately around 600,000-800,000 of oocytes in the ovary. Most of the oocytes will degenerate in the ovary. At puberty, you end up with approximately 40,000 oocytes in the ovary. Every month, a small group (15-20) of oocytes will go through this process that I just described of maturation to give rise to an ovulatory follicle here. So you start with a very large number of oocytes that will get progressively reduced to this small number approximately 500 will go through this process of ovulation. You do not need to know the

number just for general information. You start with essentially a large number of oocytes; most of the oocytes will degenerate in fact. Its not a very efficient system if you think about it. Those oogonia that are found in birth are stuck in the diplotene stage. They wont go through the process of ovulation sometimes until very late in life. So some of them are present in the ovary for over 35-40 years; so its a long time. Thats why its believed that a late pregnancy in life gives rise to more genetic problems than early in life. Under the influence of luteinizing hormone (FSH/LH), meiosis I is completed but the admission of the first polar body. So thats the first time you have haploid complement of chromosomes of this oocyte, which is known at this stage as the secondary oocyte. This oocyte will arrest in metaphase when ovulation takes place. Meiosis II is only completed and you have the admission of the 2nd polar body only upon fertilization. Slide 22: Spermatogenesis So whats happening in the male in terms of spermatogenesis, the process by which spermatogonia are transformed into spermatozoa. That begins at puberty and continues into old age. Meiosis and sperm maturation beings at puberty in the seminiferous tubules. This is a representation here of the seminiferous tubules. Along the basement membrane of the seminiferous tubules, you find the spermatogonial stem cell, the cell population that are still diploid population which is essentially a spermatogonial stem cell population that continue to proliferate throughout life. Those primary spermatocytes will divide during the first meiosis to give rise to secondary spermatocyte which thought to have haploid complement of chromosomes and give rise through the second meiosis to the spermatids, which are also haploid complement of chromosomes. Slide 23: Spermiogenesis So the differentiation of spermatids, which is here (refer to slide22), into mature sperm is a process known as spermiogenesis, the transformation of spermatids into sperm. That involves a lot of remodeling of the cell as you can seeyou go from a cell that is spherical like this to a mature sperm that has a head, middle piece and a tail. So there is a lot of reconstruction taking place here. One of them being the condensation of the nucleus and the formation of the acrosome, which covers of the nucleus and accumulate proteolytic enzymes that will be essential for fertilization of the egg and the penetration of the zona pellucida. And also there will be a resorption of most of the cytoplasm of the cell that will be completely digested by the satellite cells that are surrounding. Mature in the sense the ability to fertilize an egg. The sperm that you find in ejaculate is immature. It needs to go through the process of capacitation by travelling through the female genital tract. Slide 24: Key differences in male & female gametogenesis During oogenesis in females, you have a determined and finite population of oocytes which is established at birth. In the male, you have the continuous dividing stem cell population in the seminiferous tubules. Only one gamete is produced by each meiosis event whereas four gametes are produced in males. Meiosis begins during fetal development and is not resumed until puberty. Meiosis in male is initiated at puberty. In female, the differentiation of the gamete occurs while its still diploid, the oogonia. The gametes differentiate in male while it is haploid going through the process of spermiogenesis.

Slide 25: Fertilization So now we have the gametes that are formed. The next step (initial step) in the development of an embryo is fertilization. The fertilization is defined as the fusion between the haploid gametes, egg and sperm. It results in the formation of the diploid embryo and this is referred to as the beginning of embryogenesis. Slide 26: Fertilization takes place in the so called ampullary region of the fallopian tube. Here you have the uterus in the fallopian tube, the ovaries here. The fertilization takes place in thewhich is the broader region of the fallopian tube here. Slide 27: Ovulated oocyte The ovulated oocyte, and we talked about that, has this structure. This is the oocyte itself. I guess the nucleus is here. You have the zona pellucida that surround the oocyte and you have the first polar body here. So the second set of chromosomes that will be extruded during meiosis II is only happening during the process of fertilization when the sperm enters the egg. So thats the completion of the second meiosis Slide 28: Key features of fertilization Im going back to your point earlier. The sperm needs to go through a process of capacitation which is maturation essentially of the sperm. Sperm that has just been delivered in ejaculation has no potential to fertilize an egg. It needs to be conditioned into the female reproductive tract and this is influenced from the uterus, from the cervix, from the fallopian tubes that have interactions between the sperm and different glycoproteins that are found in the different regions of the reproductive tract that will be essential to condition the sperm. This process of conditionining of capacitation of the sperm takes approximately seven hours. The second step is contact and recognition. The binding to species-specific receptors on the zona pellucida. One of the major proteins that is found in this region are known as thethere are a number of themone of them is ZP3 which is zona pellucida protein 3 that is critical in humans for the recognition between egg and sperm. Third step is the acrosome reaction which is the release of the proteolytic enzymes that will be essential for digesting the zona pellucida to allow the sperm to enter the egg. So those proteolytic enzymes that I mentioned earlier accumulate in the acrosome, the most anterior part of the sperm, during the process of spermiogenesis. Fourth step is the fusion with the egg membrane. There is a very important process taking place here which is the cortical granule reaction of the egg, which is essentiallyand Im going to show you a slide to illustratevery important step to block polyspermy (to prevent any other sperm to enter the egg only 1 sperm can enter an oocyte at a time). So once the first sperm enters the egg, these complete block to sperm. The fifth step is the egg activation which is the completion of the meiosis (second meiotic division extrusion of the 2nd polar body). The sixth step is the fusion of the genetic material which is reconstitution of the diploid complement of chromosomes Slide 29: The sperm needs to go through capacitation, which is conditioning of the sperm to be able to enter zona pellucida here and eventually the entire head and tail will penetrate the oocyte.

Slide 30: Cortical granule reaction So the step that I mentioned which is very critical to prevent any additional sperm to enter the oocytebecause you can imagine if multiple sperm with their own chromosome complement enter the oocyte, youll have an imbalance in the chromosome composition of the oocyte. So its very important to block/prevent any other sperm to enter the oocyte. So when the sperm comes into contact with the zona pellucida, the acrosome reaction takes place which allows for the proteolytic enzymes to digest the zona pellucida and come in contact with the plasma membrane of the oocyte. As soon as the sperm comes into contact with the plasma membrane of the oocyte, there is a release of cortical granules. So the cortex of the oocyte is lined with granules that contain a number of enzymes that would be release as soon as the sperm touch the membrane of the oocyte and that will create a separation between the zona pellucida and the plasma membrane of the oocyte. And also those enzymes that released from the cortical granules will change the properties of the zona pellucida in such a way that it will not recognized by any additional sperm. Slide 31: Zygote So at the end of fertilization, you end up with a fertilized egg that looks like this. So you have the egg here with two pronuclei that are starting to fuse . Zona pellucida is still there.and you have the polar bodies that are present here, which contain the haploid complement of chromosomes. And those polar bodies have no purpose, they will eventually degenerate. Slide 32: Cleavage So once you form the zygote, the next step is the phase of cleavage, which is the division of the egg through mitotic division to give rise to 2 cells then 4 cells then 8 cells and those cells at that stage are known as blastoomeres Slide 33: Embryonic Genome Activation During those early steps that follow fertilization around the 2-4 cell stage, there is a very important step taking place which is referred to as the embryonic genome activation. Here is the unfertilized egg, fertilized egg, 2 cell stage4 cell8 cell. Here we are looking at transcription of gene that takes place at those different steps. As you can see here, the oocyte up to 1 cell stagemost of the development of the oocyte will rely on accumulation of RNA that are coming from the mother, which are known as maternal mRNA. Following fertilization, there is a sharp decrease in the accumulation of the maternal mRNA that start to be degraded. In concomitant to that process, there is a phase of zygotic genome activation that takes place around the 2-4 cell stage. So at this stage when the two pronuclei fuse, the egg for further development will not rely on maternal mRNA that been accumulated but rely on new transcription taking place at that stage and this is essential to regulate all the subsequent development of the embryo. This is a very limited burst of transcription that takes place very early on between the 2 and 8 cell stage. That will be enough, this first burst, to allow for the early development of embryo up to implantation at least. Slide 34: Compaction So when the egg goes through the mitotic division (2 cells4cell8cells), those cells initially are very loosely arranged and they have this morula type of shape. Around the 8 cell stage, the embryo will go through a step which is known as the process of compaction where the blastomere will start to maximize the contact between each other.

Slide 35: Cellular events associated with compaction This is a process that takes place at the 8 cell stage. This is mediated by an increase in tight junction formation. There is also an increase in the production of cell-cell adhesion molecule such as E-cadherin. There is formation of gap junctions. Also associated with that remodeling of the morula into a compacted egg, there is a change in the shape of cells that we will adopt this wedge shaped. There will be an establishment of a polarity. There will be cells that are more towards the outside and cells towards the inside. This polarity predicts the polarity further later the polarity of the embryo itself. Slide 36: Going back here to this diagram, fertilization takes place here in this ampullary region and then the egg will progress slowly through the fallopian tubes. Compaction takes place around the 3rd day of gestation. Slide 37: Blastocyste formation and implantation Following compaction, there is the process of blastocyst formation, which is essentially the formation of a cavity in the embryo by accumulation of liquid that is produced by the cells that are known as the trophectoderm cells. So I mentioned during the process of compaction, there will be cells more towards the outside and cells towards the inside. Cells that are more towards the outside will give rise to this outer layer which are known as the trophectoderm. Cells that more towards the inside will give rise to the cells known as the inner cell mass (ICM). ICM itself will give rise to the embryo in its entirety. The trophectoderm and trophoblast cells here that are found all around including around the blastocyst cavity and around ICM will contribute to the placenta. Slide 38: CDX2 and OCT4 expression So molecularly, we can identify those two populations of cells, the one that will contribute to the embryo and the one that will contribute to the trophectoderm or the placenta by looking at expression of specific genes that allow for identification of those two cell populations. This is a section through blastocyst. We see that those cells that are surrounding the cavity and surrounding the group of cells express the transcription factor known as CDX2 whereas those cells that are forming the ICM that will give rise to the embryo proper express this other gene, which is known as OCT4. So the expression of those two genes, I believe to be able to help make the decision between the cells that will become trophectoderm versus a cell that becomes ICM. Slide 39: Unique properties of the inner cell mass (ICM) Those cells of the ICM have very interesting properties. They are a source of embryonic stem cells (ESCs). They can self-renew if you culture them in-vitro. They are pluripotent in the sense they can give rise to any of the structure of the body that are derived from any of the 3 germ layers. Slide 40: Hatching from the zona pellucida So weve already went through fertilization, cleavage, compaction, and blastocyst formation. The next step is important for the embryo to be released from the zona pellucida and that is through the process known as hatching. The blastocyst is released from the surrounding zona

pellucida. Why is this important? Because the blastocyst at this stage needs to implant in the uterine wall and with the zona pellucida, that will not happen. So this is a very important step and this is mediated by a number of proteolytic enzymes that are produced by trophoblastic cells of the blastocyst. Slide 41: Hatching will take place around 4-5 days after fertilization. Slide 42: Blastocyst implantation The next stop following hatching is implantation. So you get rid of the zona pellucidaits not fully dissolvedits made fragile so that the blastocyst can be released from it. So when the blastocyst is free from the zona pellucida, it has the ability to implant in the uterine wall. At that stage, around the time of implantation, these cells which are known as the inner cell mass or sometimes referred to as epiblast will segregate a population of cells that will line right underneath the ICM that are known as the hypoblast, which are shown in yellow. In addition, they start the formation of a cavity here in this region just above the ICM which is the amniotic cavity. So at this stage, you have an embryo that by the time of implantation that is referred to as a bilaminar embryonic disk. You have the epiblast and this layer of hypoblast. Slide 43: GATA6 and NANOG expression Again using molecular markers, we can identify this process of segregation of hypoblast cells from epiblast looking at number of genes. This is two genes here: GATA6 and NANOG which are differentially expressed in cells of the epiblast versus cells of the hypoblasts. GATA6 will be essential to promote the formation of hypoblastic cells, those cells that will be lining the ICM. NANOG will be essential to maintain this population of inner cell mass cells which are also referred to as epiblast. So at the time of implantation, you have the two layers, the epiblast and hypoblast. Slide 44: Derivativs of the epiblast & hypoblast The epiblast will contribute to all of the derivatives of the embryo in addition to forming the embryonic endoderm, mesoderm, and form most of the ectodermal derivatives. Hypoblast, that has segregated from the epiblastic region, will give rise to the endoderm of the yolk sac and will contribute to the extraembryonic mesoderm, which will be essential for the formation of the placenta. Slide 45: So were now around day 6. Its the time of implantation of the blastocyst at this region of the endometrium. Slide 46: 7.5-day human blastocyste Just to finish, Im going to show you a couple of view of the embryo at different times of implantation. So you have here in blue, cells of the epiblast and the amniotic cavity that is starting to form here and here you have the hypoblast. The trophectoderm cells will give rise to a population of cells that are known as syncytiotrophoblast and cytotrophoblast. The syncytiotrophoblast cells will be essential in allowing the implantation of the blastocyst into the uterine wall. Those syncytiotrophoblast that are dividing very actively and moving into this

region known as the cytotrophoblast where you have a lot of nuclei but you dont see any clear indications between individual cells. Slide 47: 7.5 day human blastocyste This is a section through a human embryo around the same time (7.5 days). Here you have the epiblast and the hypoblast right underneath. This cytotrophoblast and here the syncytiotrophoblast, which is not as well organized and you have the formation of lacunae in this syncytiotrophoblast which will be very essential for blood exchange with the mother. Slide 48: 9 days human blastocyste So thats a little bit later. Its getting more and more embedded into this endometrium. Epiblasthypoblasthere is the yolk sacand you see the formation of the lacunae that are present in the syncytiotrophoblastand this is the blood vessel from the mother here. Slide 49: 12 days human blastocyste Here the yolk sac, which is derived from this epiblast here, would give rise to the extraembryonic mesoderm, which is surrounding the embryo at this stage. Little by little, the syncytiotrophoblast and the cytotrophoblast are invading the endometrium and as they are invading the endometrium they are becoming closer to contacting the blood vessels that you have in the endometrium and eventually those blood vessels will fill up the lacunae that I mentioned earlier that you find in the syncytiotrophoblast. Slide 50: 12 day human blastocyste Heres another view of an embryo at 12 days. This is the epiblasthypoblast hereyolk sac againextraembryonic mesoderm that is surrounding the embryoand we dont see very well here but the lacunae that are starting to be filled with blood. Slide 51: 13 day human blastocyste This is the 13th daya little bit later where those cytotrophoblast start to form those primary villi here which will turn into secondary and tertiary villi. We will talk about that later. Slide 52: 13 day human blastocyste This is a view of an embryo around 13 days. Again you have the epiblastyolk sacand the hypoblast is right underneathand you see the chorionic cavity here in the amnionic cavity that is just above the epiblast.