Chimera (genetics)

A chimeric mouse with its offspring

A chimera or chimaera is a single organism (usually an animal) that is composed of two or more different populations of genetically distinct cells that originated from different zygotes involved in sexual reproduction. If the different cells have emerged from the same zygote, the organism is called a mosaic. Chimeras are formed from four parent cells (two fertilized eggs or early embryos fused together). Each population of cells keeps its own character and the resulting organism is a mixture of tissues. Chimeras are typically seen in animals; there are some reports on human chimerism.^[1] Plant chimeras are discussed separately.

This condition is either inherited, or it is acquired through the infusion of allogeneic hematopoietic cells during transplantation or transfusion. In nonidentical twins, chimerism occurs by means of blood-vessel anastomoses. The likelihood of offspring being a chimera is increased if it is created via in vitro fertilization. Chimeras can often breed, but the fertility and type of offspring depends on which cell line gave rise to the ovaries or testes; varying degrees of intersexuality may result if one set of cells is genetically female and another genetically male.

Tetragametic chimerism

Tetragametic chimerism is a form of congenital chimerism. This condition occurs through the fertilization of two separate ova by two sperm, followed by the fusion of the two at the blastocyst or zygote stages. This results in the development of an organism with intermingled cell lines. Put another way, the chimera is formed from the merging of two nonidentical twins (although a similar merging presumably occurs with identical twins, but as their DNA is almost identical, the presence would not be immediately detectable in a very early (zygote or blastocyst) phase. As such, they can be male, female, or hermaphroditic.

As the organism develops, it can come to possess organs that have different sets of chromosomes. For example, the chimera may have a liver composed of cells with one set of chromosomes and have a kidney composed of cells with a second set of chromosomes. This has occurred in humans, and at one time was thought to be extremely rare, though more recent evidence suggests that it is not as rare as previously believed.^[1]

This is particularly true for the marmoset. Recent research shows most marmosets are chimerae, sharing DNA with their fraternal twins.^[2]

Most chimerae will go through life without realizing they are chimeras. The difference in phenotypes may be subtle (e.g., having a hitchhiker's thumb and a straight thumb, eyes of slightly different colors, differential hair growth on opposite sides of the body, etc.) or completely undetectable. Chimera may also show, under a certain spectrum of UV light, distinctive marks on the back resembling that of arrow points pointing downwards from the shoulders down to the lower back; this is one expression of pigment unevenness called Blaschko's lines.^[3]

Affected persons may be identified by the finding of two populations of red cells or, if the zygotes are of opposite sex, ambiguous genitalia and hermaphroditism alone or in combination; such persons sometimes also have patchy skin, hair, or eye pigmentation (heterochromia). If the blastocysts are of opposite sex, genitals of both sexes may be formed, either ovary and testis, or combined ovotestes, in one rare form of intersexuality, a condition previously known as true hermaphroditism.

Note that the frequency of this condition does not indicate the true prevalence of chimerism. Most chimerae composed of both male and female cells probably do not have an intersex condition, as might be expected if the two cell populations were evenly blended throughout the body. Often, most or all of the cells of a single cell type will be composed of a single cell line, i.e. The blood may be composed prominently of one cell line, and the internal organs of the other cell line. Genitalia produce the hormones responsible for other sex characteristics. If the sex organs are homogeneous, the individual will not be expected to exhibit any intersex traits.

Natural chimeras are almost never detected unless they exhibit abnormalities such as male/female or hermaphrodite characteristics or uneven skin pigmentation. The most noticeable are some male tortoiseshell cats or animals with ambiguous sex organs.

The existence of chimerism is problematic for DNA testing, a fact with implications for family and criminal law. The Lydia Fairchild case, for example, was brought to court after DNA testing apparently showed that her children could not be hers. Fraud charges were filed against her and her custody of her children was challenged. The charge against her was dismissed when it became clear that Lydia was a chimera, with the matching DNA being found in her cervical tissue. Another case was that of Karen Keegan, who was also in danger of losing her children, after a DNA test for a kidney transplant seemed to show she wasn't the mother of her children.^{[1] [4]}

The tetragametic state has important implications for organ or stem-cell transplantation. Chimeras typically have immunologic tolerance to both cell lines.

Microchimerism

Main article: Microchimerism

Microchimerism is the presence of a small number of cells that are genetically distinct from those of the host individual. Apparently, this phenomenon is related to certain types of autoimmune diseases, however, the mechanisms responsible for this relationship are unclear.

"Parasitic" chimerism in anglerfish

Chimerism occurs naturally in adult Ceratioid anglerfish and is in fact a natural and essential part of their life cycle. Once a male is born, it begins its search for a female. Using strong olfactory glands, the male searches until it locates a female anglerfish. The male, less than an inch in length, bites into her skin and releases an enzyme that digests the skin of his mouth and her body, fusing the pair down to the blood-vessel level. While this attachment has become necessary for the male's survival, it will eventually consume him, as both anglerfish fuse into a single hermaphroditic individual. Sometimes in this odd ritual, more than one male will attach to a single female as a 'parasite'. They will both be consumed into the body of the larger female angler. Once fused to a female, the males will reach sexual maturity, developing large testicles as their other organs atrophy. This process allows for sperm to be in constant supply when the female produces an egg, so that the chimeric fish is able to have a greater number of offspring.^[5]

Germline chimerism

Germline chimerism occurs when the germ cells (for example, sperm and egg cells) of an organism are not genetically identical to its own. It has recently been discovered that marmosets can carry the reproductive cells of their (fraternal) twin siblings, because of placental fusion during development. (Marmosets almost always give birth to fraternal twins.) ^[2][6][7]

Human chimeras

In 1953 a human chimera was reported in the British Medical Journal. A woman was found to have blood containing two different blood types. Apparently this resulted from cells from her twin brother living in her body. ^[8]. More recently, a study found that such blood group chimerism is not rare.^[9] Another report of a human chimera was published in 1998, where a male human had some partially-developed female organs due to chimerism. He was conceived by in-vitro fertilization.^[10]

Chimeras in research

In biological research, chimeras are artificially produced by selectively transplanting embryonic cells from one organism onto the embryo of another, and allowing the resultant blastocyst to develop. Chimeras are not hybrids, which form from the fusion of gametes from two species that form a single zygote with a combined genetic makeup, or Hybridomas which, as with hybrids, result from fusion of two species' cells into a single cell and artificial propagation of this cell in the laboratory. Essentially, in a chimera, each cell is from either of the parent species, whereas in a hybrid and hybridoma, each cell is derived from both parent species. "Chimera" is a broad term and is often applied to many different mechanisms of the mixing of cells from two different species.

As with cloning, the process of creating and implanting a chimera is imprecise, with the majority of embryos spontaneously terminating. Successes, however, have led to major advancements in the field of embryology, as creating chimeras of one species with different physical traits, such as colour, has allowed researchers to trace the differentiation of embryonic cells through the formation of organ systems in the adult individual.

A major milestone in chimera experimentation occurred in 1984, when a chimeric geep was produced by combining embryos from a goat and a sheep, and survived to adulthood.^[11] The creation of the "geep" revealed several complexities to chimera development. In implanting a goat embryo for gestation in a sheep, the sheep's immune system would reject the developing goat embryo, whereas a "geep" embryo, sharing markers of immunity with both sheep and goats, was able to survive implantation in either of its parent species.

In August 2003, researchers at the Shanghai Second Medical University in China reported that they had successfully fused human skin cells and dead rabbit eggs to create the first human chimeric embryos. The embryos were allowed to develop for several days in a laboratory setting, then destroyed to harvest the resulting stem cells.^[12] In 2007, scientists at the University of Nevada School of Medicine created a sheep whose blood contained 15% human cells and 85% sheep cells.^[13] The implications of increasingly realizable projects using human-animal hybrids for biopharmaceutical production, and potentially for producing cells or organs, have raised a host of ethical and safety issues.

Mouse chimeras

Chimeric mice are important tools in biological research, as they allow the investigation of a variety of biological questions in an animal that has two distinct genetic pools within it. These include insights into such problems as the tissue specific requirements of a gene, cell lineage, and cell potential. The general methods for creating chimeric mice can be summarized either by injection or aggregation of embryonic cells from different origins. The first chimeric mouse was made by Beatrice Mintz in the 1960s through the aggregation of eight cell stage embryos.^[14] Injection on the other hand was pioneered by Richard Gardner and Ralph Brinster who injected cells into blastocysts to create chimeric mice with germ lines fully derived from injected ES Cells.^[15] Mouse embryos both periimplantation and post implantation can contribute to a chimera. It is post implantation that ES cells from the inner cell mass of a blastocyst can contribute to all cell lineages of a mouse including the germ line. ES cells are also a useful tool in chimeras because genes can be mutated in them through the use of homologous recombination, thus allowing gene targeting. Since this discovery occurred in 1999, ES cells have become a key tool in the generation of specific chimeric mice.^[16]

Underlying biology

The ability to make mouse chimeras comes from an understanding of early mouse development. Between the stages of fertilization of the egg and the implantation of a blastocyst into the uterus, different parts of the mouse embryo retain the ability to give rise to a variety of cell lineages. Once the embryo has reached the blastocyst stage, it is composed of several parts, mainly the trophectoderm, the inner cell mass, and the primitive endoderm. Each of these parts of the blastocyst gives rise to different parts of the embryo; the inner cell mass gives rise to the embryo proper, while the trophectoderm and primitive endoderm give rise to extra embryonic structures that support growth of the embryo.^[17] Two- to eight-cell-stage embryos are competent for making chimeras, since at these stages of development, the cells in the embryos are not yet committed to give rise to any particular cell lineage, and could give rise to the inner cell mass or the trophectoderm. In the case where two diploid eight-cell-stage embryos are used to make a chimera, chimersim can be later found in the epiblast, primitive, endoderm and trophectoderm of the mouse blastocyst.^[18][19] It is possible to dissect the embryo at other stages so as to accordingly give rise to one lineage of cells from an embryo selectively and not the other. For example, subsets of blastomeres can be used to give rise to chimera with specified cell lineage from one embryo. The Inner Cell Mass of a diploid blastocyst for example can be used to make a chimera with another blastocyst of eight-cell diploid embryo; the cells taken from the inner cell mass will give rise to the primitive endoderm and to the epiblast in the chimera mouse.^[20] From this knowledge, ES cell contributions to chimeras have been developed. ES cells can be used in combination with eight-cell-and two-cell-stage embryos to make chimeras and exclusively give rise to the embryo proper. Embryos that are to be used in chimeras can further be genetically altered in order to specifically contribute to only one part of chimera. An example is the chimera built off of ES cells and tetraploid embryos, tetraploid embryos which are artificially made by electrofusion of two two-cell diploid embryos. The tetraploid embryo will exclusively give rise to the trophectoderm and primitive endoderm in the chimera ^[21][22]

Methods of production

There are a variety of combinations that can give rise to a successful chimera mouse and — according to the goal of the experiment — an appropriate cell and embryo combination can be picked; they are generally but not limited to diploid embryo and ES cells, diploid embryo and diploid embryo, ES cell and tetraploid embryo, diploid embryo and tetraploid embryo, ES cells and ES cells. The combination of embryonic stem cell and diploid embryo is a common technique used for the making of chimeric mice, since gene targeting can be done in the embryonic stem cell. These kinds of chimeras can be made through either aggregation of stem cells and the diploid embryo or injection of the stem cells into the diploid embryo. If embryonic stem cells are to be used for gene targeting to make a chimera, the following procedure is common: a construct for homologous recombination for the gene targeted will be introduced into cultured mouse embryonic stem cells from the donor mouse, by way of electroporation; cells positive for the recombination event will have antibiotic resistance, provided by the insertion cassette used in the gene targeting; and be able to be positively selected for.^[23][24] ES cells with the correct targeted gene are then injected into a diploid host mouse blastocyst. These injected blastocysts are then implanted into a pseudo pregnant female surrogate mouse which will bring the embryos to term and give birth to a mouse whose germline is derived from the donor mouse's ES cells.^[25] This same procedure can be achieved through aggregation of ES cells and diploid embryos, diploid embryos are cultured in aggregation plates in wells were single embryos can fit, to these wells ES cells are added the aggregates are cultured until a single embryo is formed and has progressed to the blastocyst stage, and can then be transferred to the surrogate mouse.^[26]

Legislation

The US and Western Europe have strict codes of ethics and regulations in place that expressly forbid certain subsets of experimentation using human cells, though there is a vast difference in the regulatory framework.^[27] In May 2008, a robust debate in the House of Commons of the United Kingdom on the ethics of creating chimeras with human stem cells led to the decision that embryos would be allowed to be made in laboratories, given that they would be destroyed within the first 14 days. No such foundation has been set for chimera research regulation in the US.

See also

• Blaschko's lines
• Chimera (plant)
• Chimerism in human intersexuality
• Genetic chimerism in fiction
• Lydia Fairchild
• Mosaic (genetics)
• Parahumans
• Vanishing twin
• Conjoined twins

References

1. ^ ^{a b c} Norton, Aaron; Ozzie Zehner (2008). "Which Half Is Mommy?: Tetragametic Chimerism and Trans-Subjectivity". Women's Studies Quarterly Fall/Winter: 106–127. http://ucdavis.academia.edu/AaronTNorton/Papers/126923/Which_Half_Is_Mommy_Tetragametic_Chimerism_and_Trans-Subjectivity. 
2. ^ ^{a b} Ross, C. N.; J. A. French, G. Orti (2007). "Germ-line chimerism and paternal care in marmosets (Callithrix kuhlii)". Proceedings of the National Academy of Sciences 104 (15): 6278–6282. doi:10.1073/pnas.0607426104. ISSN 0027-8424. PMC 1851065. PMID 17389380. http://www.pnas.org/cgi/content/abstract/0607426104v1. 
3. ^ Dr. Barry Starr. "2) In the chimera episode, the fact that the guy was a chimera was first established through a camera flash on the skin illuminating skin anomalies...". Ask a Geneticist. Stanford School of Medicine. http://www.thetech.org/genetics/ask.php?id=75. 
4. ^ "The Twin Inside Me: Extraordinary People". Channel 5 TV, UK. 23:00 9 March 2006. Archived from the original on May 26, 2006. http://web.archive.org/web/20060526105634/http://www.five.tv/programmes/extraordinarypeople/twininside/. 
5. ^ http://en.wikipedia.org/wiki/Ceratiidae
6. ^ Zimmer, Carl (2007-03-27). "In the Marmoset Family, Things Really Do Appear to Be All Relative". The New York Times. http://www.nytimes.com/2007/03/27/science/27marm.html. Retrieved 2010-04-01. 
7. ^ Hooper, Rowan (26 March 2007). "Marmosets may carry their sibling's sex cells". New Scientist. http://www.newscientist.com/article/dn11464-marmosets-may-carry-their-siblings-sex-cells.html. 
8. ^ Bowley, C. C.; Ann M. Hutchison, Joan S. Thompson, Ruth Sanger (July 11 1953). "A human blood-group chimera". British Medical Journal: 81. http://www.bmj.com/content/2/4827/81.full.pdf. Retrieved 20111002. 
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10. ^ Strain, Lisa; John C.S. Dean, Mark P. R. Hamilton, David T. Bonthron (1998). "A True Hermaphrodite Chimera Resulting from Embryo Amalgamation after in Vitro Fertilization". The New England Journal of Medicine 338: 166–169. http://www.nejm.org/doi/full/10.1056/NEJM199801153380305. 
11. ^ "It's a Geep". Time. 27 February 1984. http://jcgi.pathfinder.com/time/archive/preview/0,10987,921546,00.html. Retrieved 2 August 2006. 
12. ^ Mott, Maryann (January 25, 2005). "Animal-Human Hybrids Spark Controversy". National Geographic News. http://news.nationalgeographic.com/news/2005/01/0125_050125_chimeras.html. 
13. ^ "Iranian scientist creates sheep with half-human organs". Press TV. 27 Mar 2007. http://classic-web.archive.org/web/20071114210217/http://www.presstv.ir/detail.aspx?id=3995&sectionid=3510208. 
14. ^ Mintz B., Silvers W. K. (1967). ""Intrinsic" Immunological Tolerance in Allophenic Mice". Science 158 (3807): 1484–6. doi:10.1126/science.158.3807.1484. PMID 6058691. 
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19. ^ Rossant, J. (1976). "Postimplantation development of blastomeres isolated from 4- and 8-cell mouse eggs". J. Embryol. Exp. Morphol 36 (2): 283–290. PMID 1033982. 
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22. ^ Nagy, A. and Rossant J. (1999) Production of Es-cell aggregation chimeras. Gene Targeting: A Practical Approach (Joyner, A, ed.) IRL Press at Oxford University, Oxford, UK
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25. ^ Chimera Mouse production by blastocyst injection, Wellcome trust Sanger Institute, http://www.eucomm.org/docs/protocols/mouse_protocol_1_Sanger.pdf
26. ^ Tanaka M, Hadjantonakis AK, Nagy A (2001). "Aggregation chimeras. Combining ES cells, diploid and tetraploid embryos.". Methods in molecular biology (Clifton, N.J.) 158: 135–54. doi:10.1385/1-59259-220-1:135. PMID 11236654. 
27. ^ Futehally, Ilmas, Beyond Biology, Strategic Foresight Group [2]

• Yu N, Kruskall MS, Yunis JJ, Knoll JH, Uhl L, Alosco S, Ohashi M, Clavijo O, Husain Z, Yunis EJ, Yunis JJ, Yunis EJ (2002). "Disputed maternity leading to identification of tetragametic chimerism". N Engl J Med 346 (20): 1545–52. doi:10.1056/NEJMoa013452. PMID 12015394. 
• Appel, Jacob M. "The Monster's Law", Genewatch, Volume 19, Number 2, March–April 2007.
• Ainsworth, Claire (November 15, 2003). The Stranger Within. New Scientist (subscription). (reprinted here)
• Nelson, J. Lee (Scientific American, February 2008). Your Cells Are My Cells
• Weiss, Rick (August 14, 2003). Cloning yields human-rabbit hybrid embryo. The Washington Post.
• Weiss, Rick (February 13, 2005). U.S. Denies Patent for a too-human hybrid. The Washington Post.
• Interesting Article with a very easy explanation of Chimera [3]
• L. M. Repas-Humpe, A. Humpe, R. Lynen, B. Glock, E. M. Dauber, G. Simson, W. R. Mayr, M. Köhler, S. Eber (1999). "A dispermic chimerism in a 2-year-old Caucasian boy". Journal Annals of Hematology 78 (9): 431–434. doi:10.1007/s002770050543. 
• Strain, Lisa; Dean, John C.S.; Hamilton, Mark P.R.; Bonthron, David T. (1998). "A True Hermaphrodite Chimera Resulting from Embryo Amalgamation after in Vitro Fertilization". New England Journal of Medicine 338 (3): 166–9. doi:10.1056/NEJM199801153380305. PMID 9428825. 
• Chimerism and cellular mosaicism, Genetic Home Reference, U.S. National Library of Medicine, National Institute of Health.