In vitro fertilization
This will be a disorganized mess for a bit.
- in vitro fertilization
- embryo, sperm, oocyte/egg cryopreservation
- embryo microinjection (CRISPR, other genetic payloads, etc) (but really should genetically modify upstream cells instead of embryo-stage, or modify embryonic stem cell cultures, etc...)
- intracytoplasmic sperm injection, spermatid injection, (spermatocyte injection?)
- in vitro gametogenesis, spermatogenesis, oogenesis/oocyte production
- embryo splitting (blastocyst bisection, blastomere separation)
- embryo culture media/ATP delivery for infinity embryo splitting
- interspecies blastocyst complementation (tweet thread; logs)
- xenogeneic stem cell transplantation
- organogenic niche method
- interspecies pregnancy, xenogenic embryo, xenotransplantation; co-gestation of interspecies embryos.
- interspecies trophoblast embryo envelopment
- somatic cell nuclear transfer, various cloning techniques
- somatic cell hybridization, somatic cell haploidization
- somatic cell fusion, other cell electrofusion
- inner cell mass transfer
- Velocimouse technique
- interspecies nuclear transfer, cytoplasm transfer, etc
- gestation using ex vivo uterus (not the same as an artificial womb)
- uterus transplantation (works for human pregnancy), uterus xenotransplantation, uterus development by blastocyst complementation
- ovary follicle transplantation, ovary transplantation (into the arm), ovary cryopreservation
- oocyte in vitro maturation (ooIVM)
- embryonic stem cell extraction and culture, expansion
- "Non-embryo-destructive extraction of pluripotent embryonic stem cells" https://www.thieme-connect.com/products/ejournals/html/10.1055/s-0035-1558183
- host/child microchimerism
- germ cell transplantation
- xenografting of testicular tissue pieces for sperm generation
- adult sperm stem cell gene therapy; testis-mediated gene therapy; testis electroporation.
- pronuclear injection
- ooplasm transfer, ooplasm xenotransfer, interspecies ooplasm transfer
- xy/xx sperm sorting (xyinc: logs)
- human cloning is a reasonable pathway to fixing human infertility beyond the normal reproductive age
- we really ought to do more GWAS and genetics studies of individuals or families that experience exceptionally easy, quick, or painless childbirths, or other pregnancy related studies (morning sickness has been studied at least, and some mutations are known there)
Sperm sorting for sex selection
Sperm sorting for sex selection primarily relies on flow cytometry combined with fluorescent DNA staining, a technique commercialized as MicroSort and based on the Beltsville Sperm Sexing Technology. The method exploits the ~2-4% DNA content difference between X-bearing and Y-bearing sperm (X chromosomes carry more genetic material than Y chromosomes). Sperm are stained with Hoechst 33342, a fluorescent dye that binds to DNA, causing X-bearing sperm to fluoresce slightly brighter than Y-bearing sperm due to their greater DNA content. The stained sperm are then passed single-file through a flow cytometer where a laser excites the dye and detectors measure fluorescence intensity. Based on this measurement, droplets containing individual sperm are given an electrical charge and deflected by an electromagnetic field into separate collection tubes.
"… However, the only proven method for separating male- and female-bearing sperm is the fluorescence activated cell sorting approach of Johnson et al. (1987a, 1987b, 1989, and 1999). Also known as the “Beltsville Sperm Sexing Technology,” this method was subsequently licensed to XY, Inc. (www.xyinc.com) for commercial development. …Consequently, some sex cells contain X chromosomes, and others Y chromosomes. Gametes with the X chromosome contain 4% more of DNA than sperm with the Y chromosome. While staining chromosomes of germinal cells, it was found that gametes with X chromosomes absorb 4-5% more of stain than gametes with Y chromosomes."
XY, Inc. was heavily involved in legal battles regarding this technology, and there were allegations of securities fraud involving executive Mervyn Jacobson - "Bull semen sexing battle leads to $330M award"
techniques to consider:
- MicroSort / Beltsville sperm sexing technology
- flow cytometry + DNA fluorescent labeling dye
- double sperm cloning
- density-gradient centrifugation
- swim-up / sperm washing
- albumin column separation
- magnetic-activated cell sorting with annexin-V
- spiral inertial microfluidics
- dean flow fractionation microfluidics
- microfluidic geometry to sort sperm cells (active sorting, passive sorting)
- sperm electrophoresis
- centrifugal counter-current distribution
- sperm sex-specific antigen labeling and immunological column or antibody-painted beads
- gold particle (AuNP) + cell penetrating peptides to label DNA?
- triplex hybridization probe assay
- optical interferometric measurement of subtle differences in refractive index or scattering between X‑ and Y‑sperm
- pH or chemical gradient or buffer to favor survival of certain sperm
- atomic force microscopy surface tomography mapping or classification
- upstream spermatogenesis modifications like chromosome duplications, or sex ratio bias via genetic intervention in sperm stem cells
- genetic engineering - sex-linked fluorescent reporters in sperm ells
- programmed cell death (apoptosis) for sperm cells with the wrong chromosome
- design a selection experiment for bacteria or other microbes that are able to selectively kill sperm that do not have an X chromosome or do not have a Y chromosome
- aptamer sperm selection
- quantitative phase imaging (QPI)
- impedance cytometry
- maybe just use optical microscope + deep learning to classify sperm cells based on surface features
- nanopore sorting technique with electrical measurement
random
Rules of motor innervation in chick embryos with supernumerary limbs (1981)
Application of the quail-chick chimera system to the study of brain development and behavior
"Hatched chicks with chimeric brains containing cells from both the domestic chicken (Gallus gallus domesticus) and the Japanese quail (Coturnix coturnix japonica) have been produced by transplantation of various regions of the neural tube at the 8- to 15- somite stage. The positions of host and donor cells relative to graft boundaries observed throughout embryonic development and after hatching implicated both radial and tangential cell movements in brain morphogenesis. In addition, transplants containing the entire quail mesencephalon and diencephalon resulted in the transfer of certain aspects of species-typical crowing behavior."
interspecies blastocyst complementation for interspecies liver organ
embryonic face transplant via fetal surgery
moving or transplanting limb bud grafts between embryos
more
hybrids, mosaics, chimeras and chimerism:
- https://en.wikipedia.org/wiki/Humanzee ("humanzee, chuman, manpanzee or chumanzee")
- https://en.wikipedia.org/wiki/Humster
"experimentally replicating the bioelectric characteristics of the site of eye formation in other parts of the body of frog embryos via microinjection of ion channel mRNA can cause eye formation in ectopic regions" (ref)
time-displanced clones and twins: https://arxiv.org/abs/1301.4299
Blastocyst complementation: current progress and future directions in xenogeneic organogenesis
polygenic embryo scoring (PGT), bostromian iterated embryo selection, direct embryo/stem cell genetic engineering
logs:
- earlier stuff in the logs not yet documented
- https://gnusha.org/logs/2023-01-02.log, including surgical embryo engineering vs genetic engineering to achieve morphological bodyplan outcomes, human-chimpanzee hybrids, xenobots, etc.
- https://gnusha.org/logs/2023-01-05.log (interspecies blastocyst complementation)
- https://gnusha.org/logs/2023-01-21.log (xy/xx sperm sorting technology)
- https://gnusha.org/logs/2023-02-08.log (interspecies pregnancy)
- https://gnusha.org/logs/2023-02-09.log (more interspecies pregnancy, embryo twinning)
library: https://diyhpl.us/~bryan/papers2/bio/in-vitro-fertilization/?C=M;O=D
additional IVF-related research notes: https://diyhpl.us/~bryan/papers2/bio/in-vitro-fertilization/notes.txt
See https://gattacastack.com/ for an index of IVF related companies focusing on human embryo editing, artificial wombs, ?embryo selection, embryo intelligence/IQ scoring, etc.
See also cloning, xenotransplantation