Jan Evangelista Purkyne, also called Johannes or Johann Evangelist Purkinje, studied cells in the cerebellum, fibers of the heart, subjective visual phenomenon, and germinal vesicle, in eastern Europe during the early nineteenth century. His investigations provided insights into various mechanisms and structures of the human body. Purkyne introduced techniques for decalcification of bones and teeth, embedding of tissue specimens, and eye examinations. He was one of the first to adopt the microtome in his experiments and to use the term protoplasm when describing the contents of young animal embryos. Purkyne identified structures in the eggs of chickens, such as the germinal vesicle, from which he hypothesized the female reproductive cell (ovum) developed, and around which he said an embryo developed. Purkyne's results helped others locate ova in mammals.
In a series of experiments between 1960 and 1965, Robert Geoffrey Edwards discovered how to make mammalian egg cells, or oocytes, mature outside of a female's body. Edwards, working at several research institutions in the UK during this period, studied in vitro fertilization (IVF) methods. He measured the conditions and timings for in vitro (out of the body) maturation of oocytes from diverse mammals including mice, rats, hamsters, pigs, cows, sheep, and rhesus monkeys, as well as humans. By 1965, he manipulated the maturation of mammalian oocytes in vitro, and discovered that the maturation process took about the same amount of time as maturation in the body, called in vivo. The timing of human oocyte maturation in vivo, extrapolated from Edwards's in vitro study, helped researchers calculate the timing for surgical removal of human eggs for IVF.
In 2009, Shoukhrat Mitalipov, Masahito Tachibana, and their team of researchers developed the technology of mitochondrial gene replacement therapy to prevent the transmission of a mitochondrial disease from mother to offspring in primates. Mitochondria contain some of the body's genetic material, called mitochondrial DNA. Occasionally, the mitochondrial DNA possesses mutations. Mitalipov and Tachibana, researchers at the Oregon National Primate Research Center in Beaverton, Oregon, developed a technique to remove the nucleus of the mother and place it in a donor oocyte, or immature egg cell, with healthy mitochondria. The resulting offspring contain the genetic material of three separate individuals and do not have the disease. Mitalipov and Tachibana's technology of mitochondrial gene replacement built on decades of research by different scientists and enables researchers to prevent the transmission of human mitochondrial diseases from mother to offspring.
Shoukhrat Mitalipov, Masahito Tachibana, and their team of researchers replaced the mitochondrial genes of primate embryonic stem cells via spindle transfer. Spindle replacement, also called spindle transfer, is the process of removing the genetic material found in the nucleus of one egg cell, or oocyte, and placing it in another egg that had its nucleus removed. Mitochondria are organelles found in all cells and contain some of the cell’s genetic material. Mutations in the mitochondrial DNA can lead to neurodegenerative and muscle diseases. Mitalipov and Tachibana used spindle replacement to produce healthy offspring from an egg with mutated mitochondria in rhesus macaques (Macaca mulatta). The experiment showed that spindle transfer eliminated the chance of transmission of mitochondrial diseases from the affected primates to their offspring, offering the potential to eliminate mitochondrial diseases in humans.
Robert Geoffrey Edwards, a British developmental biologist at University of Cambridge, began exploring human in vitro fertilization (IVF) as a way to treat infertility in 1960. After successfully overcoming the problem of making mammalian oocytes mature in vitro in 1965, Edwards began to experiment with fertilizing matured eggs in vitro. Collaborating with other researchers, Edwards eventually fertilized a human egg in vitro in 1969. This was a huge step towards establishing human IVF as a viable fertility treatment. During the four years in which Edwards experimented with IVF, he experienced many setbacks. These failures in fertilizing oocytes in vitro, however, contributed to the understanding of how fertilization did or did not happen, which was sometimes different from established dogmas. Edwards also collaborated with gynecologist and surgeon Patrick Christopher Steptoe to study sperm capacitation, which became the overture that heralded a series of successes for the team, culminating in the generation of the first test-tube baby Louise Joy Brown in 1978.