Reproductive BioMedicine Online (2012) 25, 118– 127

www.sciencedirect.com
www.rbmonline.com

REVIEW

IVF and embryo transfer: historical origin

and development
John D Biggers

Department of Cell Biology, 240 Longwood Avenue, Harvard Medical School, Boston, MA 02115, USA
E-mail address: john_biggers@hms.harvard.edu

John Biggers, DSc, PhD is professor of cell biology at Harvard Medical School. His current research interests are
evaporative drying of spermatozoa, vitrification, embryo culture, embryo assessment and the biography of
Walter Heape. He is a former Commonwealth Fellow of St John’s College, Cambridge, past President of the
Society of Reproduction, former Editor in Chief Biology of Reproduction, Chief Scientific Advisor to the Ethics
Committee, US Department of Health, Education and Welfare that made recommendations on IVF and embryo
transfer, Fellow of the American Association for the Advancement of Science, Hartman Award of the Society of
Reproduction, Pioneer Award of the International Embryo Transfer Society, Marshall Medal of the Society for the
Study of Fertility and a Life Member of the New England Fertility Society and the Society for the Study of
Reproduction.

Abstract IVF and embryo transfer for the treatment of human infertility has now resulted in the birth of over 4 million babies. The
technique did not arise as a quantum event but was built on the efforts of many earlier workers in the fields of reproductive endo-
crinology and development. One should remember the famous saying of Isaac Newton: ‘If I have seen further than most, it is because
I have stood on the shoulder’s of giants’. Ethical and moral issues have always arisen when investigators study early mammalian
development, particularly human development. This paper documents these earlier studies and also draws attention to the ethical
and moral arguments that inevitably arose. RBMOnline
ª 2012, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.
KEYWORDS: ethical issues, history, in-vitro fertilization, IVF, Gregory Pincus, John Rock

Introduction part by Johnson (2011) and on the web (www.IVF-World-

The 2010 Nobel Prize for Physiology or Medicine was awarded
to Robert Edwards for developing IVF and embryo transfer
(IVF/ET) to treat infertility in women with non-patent ovi-
ducts. His work resulted in the birth of the first ‘test tube’
baby in July 1978. Now, after a period of about 33 years,
more than 4 million babies have been born using IVF/ET,
and a new specialty of assisted reproduction has been estab-
lished with its own professional societies. The history of
IVF/ET is extensive and it has been recently documented in
wide.com). The technique did not arise as a quantum event
but was built on the efforts of many earlier workers in the
fields of reproductive endocrinology and development. One
should remember the famous quotation from Isaac Newton:
‘If I have seen further than most, it is because I have stood on
the shoulders of giants’. Ethical and moral issues have
always arisen when investigators study early mammalian
development, particularly human development. This paper
documents these early studies and also draws attention to
the ethical and moral arguments that inevitably arose.

1472-6483/$ - see front matter ª 2012, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.rbmo.2012.04.011
History of IVF and embryo transfer
IVF/ET in mythology
The idea of transferring a human fetus from one mother to
another can be found in a story from the Jain religion about
the religious leader Mahavira. The following account is a
brief summary of the full story in the Sacred Books of the
East (Jaina Sutra, 1964). At one of his reincarnations the
ruler of all the gods of heaven and earth, Sakra, realized
that the Mahavira had been conceived by Devananda, a
woman of an unacceptable caste. Sakra then commanded
that the embryo carried by Devananda be transferred into
the womb of Trisala, a woman of an acceptable caste,
and that Trisala’s embryo be placed in Devananda’s womb.
The story has been memorialized at least until the 15th cen-
tury in sculpture and painting.
Treatment of non-patent oviducts prior to
IVF/ET
Clinicians knew, by the middle of the 19th century, that
blocked oviducts resulted in sterility (Churchill, 1846).
Demands for treatments were driven by the effects of infer-
tility on the dispersal of family wealth, and since that time
several surgical procedures have been tried (reviewed by
Biggers, 1984). In 1849, Tyler Smith attempted to unblock
the oviduct in a patient by passing a whalebone bougie
through the tube (Smith, 1849). Colleagues greeted the pro-
cedure with skepticism and it was never adopted. The dem-
onstration at the end of the 19th century that the ovaries
could be transplanted to ectopic sites paved the way for
the discovery of the ovarian hormones. The fact that the
ovary maintains its function when placed in an ectopic site
led Morris (1895), an American gynaecologist, to treat
patients with blocked oviducts by grafting ovarian tissue
into the oviduct or uterus below the obstruction. The
method was never successful. In 1909, Estes, another Amer-
ican gynaecologist, introduced an operation in which the
ovary was inserted through the uterine wall, keeping its
pedicle containing blood vessels and nerves intact, where
it was hoped ovulation would occur (Estes, 1909). The
so-called Estes operation was seldom successful, although
it was used until the middle of the 20th century. While
IVF/ET was being developed in the 1970s, microsurgery
was attracting the attention of gynaecologists for the anas-
tomosis of the cut ends of the oviduct after removal of an
obstruction. By the time the first baby was produced by
IVF/ET in 1978, microsurgery procedures showed promise
of being an alternative procedure (Eddy, 1981; Winston,
1981). Since then other improvements in technique have
occurred so that the suggestion has been made that IVF/ET
and surgery should be regarded as complimentary tech-
niques for the treatment of tubal infertility (Gomel and
McComb, 2006; Schippert et al., 2010a,b).
Basic research that paved the way to current
reproductive technologies
IVF/ET involves four main aspects: (i) acquisition in suffi-
cient numbers of meiotically and cytoplasmically mature
ova; (ii) fertilization of these mature ova in vitro; (iii)
culture of preimplantation embryos; and (iv) embryo
119
replacement within a mother. Initially, these areas were
largely studied independently and it was only later that they
were linked to create the technical procedure called
IVF/ET. Unfortunately, there is a tendency in historical
accounts of IVF/ET, particularly among clinicians, to focus
almost exclusively on the actual process of fertilization
in vitro, ignoring the history of the other three components.
Walter Heape reported in 1891 that he had successfully
transferred mammalian (rabbit) embryos from one mother
to another (Heape, 1891). He was a gentleman scientist
who studied under Francis Balfour in Cambridge. Although
he never earned a degree, he was later appointed a demon-
strator in the embryology course where students recovered
and observed living rabbit embryos (Foster and Balfour,
1883). Heape transferred two ova from an Angora doe rabbit
into the Fallopian tube of a Belgian Hare recipient and
obtained six young; two had Angora phenotypes and four
had Belgian hare phenotypes.
IVF was first attempted using rabbits and guinea pigs by
Schenk (1887) at a time when the essential biological fea-
tures of fertilization were being worked out independently
by Van Beneden, Hertwig and Folin in rabbits, sea urchins
and starfish (reviewed by Austin, 1961). Another early
attempt was made by Onanoff (1893) using rabbits.
Schenk’s experiments failed and Onanoff’s results were
unconvincing.
The first successful attempts to culture preimplantation
mammalian embryos were made in 1913 by Albert Brachet,
Director of the Brussels School of Embryology at the War-
ocqué Institute of Anatomy (Brachet, 1913). He studied
the expansion of the rabbit blastocyst in vitro. His experi-
ments were done only 4 years after the first report of tissue
culture in which nerve cells were grown by Harrison at Yale
University in 1907 (Harrison, 1907). Little attention, if any,
was paid to the manipulation of mammalian development
in vitro during the next 18 years other than science fiction
accounts of ectogenesis – the complete production of an
individual outside an organism’s body. Ectogenesis was first
invoked in a lecture given in 1923 by the geneticist and bio-
chemist JBS Haldane to the Heretics Society at the Univer-
sity of Cambridge. The main objective of this society was
to needle religious dogma. Haldane made the tongue in
cheek prediction that ectogenesis would be perfected by
1960. He imagined what a student might write in the 1960s,
as follows:
It was 1951 that Dupont and Schwartz produced the first
ectogenetic child . . . France was the first country to
adopt ectogenesis officially, and by 1968 was producing
60,000 children annually by this method. In most coun-
tries the opposition was far stronger, and was intensified
by the Papal Bull ‘Nunquam prius audito’, and by the sim-
ilar ‘fetwa’ of the Khalif, both of which appeared in
1960. (Haldane, 1923)
A few years later, the notion of ectogenesis became
widely disseminated by the book by Aldous Huxley called
Brave New World, published in 1932 (Huxley, 1932). The
techniques imagined by Huxley were remarkably realistic.
Elsewhere I have speculated that his ideas may have
resulted from conversations with Gregory Pincus, who was
doing research on fertilization and development using rab-
bits in Cambridge at the time (Biggers, 1991).
120
The contributions of Gregory Pincus in this field were
highly significant, but they have been frequently overlooked
because of his major contributions to the development of
the oral contraceptive pill (Ingle, 1971; Speroff, 2009).
After obtaining his Doctor of Science degree in the Depart-
ment of Biology, Harvard University in 1927, Pincus was
awarded a National Research Council Fellowship for 3 years,
the first two at Harvard and the third at the University of
Cambridge in England under the reproductive biologist John
Hammond and at the Kaiser Wilhelm Institute in Berlin
under the geneticist R Goldschmidt. He was appointed Assis-
tant Professor of Biology in 1931, soon after his return to
Harvard, where he remained for 6 years. His first paper in
reproductive biology was entitled ‘Observations on the
living eggs of the rabbit’ (Pincus, 1930). In it he described
the work he had done in Cambridge, England, which
included a brief description of his first unsuccessful
attempts to fertilize rabbit ova in vitro. Three important
papers in the field were written during the 6 years he was
at Harvard. In 1934, he and Enzmann reported in Proceed-
ings of the National Academy of Sciences, USA that they
had successfully produced newborn rabbits following IVF.
These results were accepted by the scientific community
for many years as the first demonstration of IVF. They were
not challenged, as shall be discussed, until the 1950s when
further information on the nature of fertilization became
available. The results also attracted comments in the popu-
lar press, as happens so often with topics in reproductive
biology. The results were well received by some commenta-
tors, who suggested they may eventually help in the solution
of human problems. Others were critical, saying that the
scientists were playing God. A year later, in 1935, Pincus
and Enzmann showed that when rabbit oocytes were iso-
lated from the Graafian follicle and placed in culture they
would resume meiosis spontaneously, passing from the
arrested dictyate stage to the metaphase-II stage. Four
years later, in 1939, Pincus and Saunders demonstrated that
isolated human oocytes would also complete meiosis
in vitro, although they underestimated the duration possibly
by using partially matured oocytes. This phenomenon is
exploited in current IVF protocols. In 1936, Pincus published
a monograph entitled ‘The eggs of mammals’ with a chapter
on the culture of the initial stages of mammalian develop-
ment. He used methods practised at the time in the field
of tissue culture that used media based on biological fluids,
such as blood serum. Also, in 1936, Pincus and Enzmann
reported that they had successfully activated rabbit ova,
causing them to begin the cleavage divisions (Pincus and
Enzmann, 1936). This paper attracted the popular press
and attendant adverse publicity. WL Laurence, writing in
New York Times on 27 March 1936, speculated on some of
the possible consequences of the research Pincus had done
over the previous few years as follows:
As rabbits and men belong to the mammalian group, the
work is viewed as pointing toward the possibility of
human children being brought into the world by a
‘host-mother’ not related by blood to the child.
It is reasoned that eventually women capable of having
children whose health does not permit them to do so
may ‘hire’ other women to bear their children for them,
children actually their own flesh and blood.
JD Biggers
To one who desires to speculate at this point the Harvard
experiment offers another possibility. Theoretically, at
least, it may become possible for a woman so inclined,
particularly in a country influenced by eugenic
considerations, to bring into the world twelve children
a year by ‘hiring’ twelve ‘host-mothers’ to bear their
test-tube-conceived children for them.
Advocates of ‘race betterment’ might urge such proce-
dures for men and women of special aptitudes, physical,
mental or spiritual (Laurence, 1936).
The following day, an emotionally negative editorial was
published in the New York Times entitled ‘Brave new
world’. A few months later, JD Radcliff, wrote a sensational
article for Collier’s Magazine with the title ‘No father to
guide them’. Radcliff’s article contained an unflattering
photograph of Pincus smoking a cigarette holding two adult
rabbits that had not been produced parthenogenetically,
and commented:
In the resulting world man’s value would shrink. It is con-
ceivable that the process would not even produce males.
The mythical land of the Amazons would then come to
life. A world where woman would be self-sufficient;
man’s value precisely zero.
Radcliff recorded that Pincus found such comments
about his work annoying saying ‘. . . I am not interested in
the implications of the work’ (Radcliff, 1937).
In 1937, Pincus was granted sabbatical leave by Harvard
to spend another year in Cambridge, UK, but was informed
that his assistant professorship would not be renewed on his
return to Harvard. Thus, when he returned to the USA he
was unemployed. The sensational publicity may have been
a cause of Pincus’s losing his faculty appointment. Many
conservative academics probably believed that it was a
threat to Harvard’s image. However, there were other
causes. The President of Harvard, James B Conant, replaced
Pincus’s mentor, the Chairman of the Department of Biol-
ogy, with someone more oriented to a molecular approach
to the subject (Speroff, 2009).
1937 was also a year when new ideas changed clinical
approaches to infertility. In 1937 the following anonymous
editorial was published in New England Journal of
Medicine:
Conception in a watch glass
Contemplating this new discovery, one’s mind travels
much farther. Lewis and Hartman have isolated a fertil-
ized monkey ovum and photographed its early cleavage
in vitro. Pincus and Enzmann have started one step ear-
lier with the rabbit, isolating an ovum, fertilizing it in a
watch glass, and re-implanting it in a doe other than
the one that furnished the egg, and have thus success-
fully inaugurated pregnancy in an unmated animal. If
such an accomplishment with rabbits were to be dupli-
cated in human beings, we should, in the words of ‘flam-
ing youth’, be ‘going places’. The difficulty with human
ova has been that those recovered from tubes have
regressed beyond the possibility of fertilization
in vitro. But by utilizing the electrical sign we may be
able to obtain them from the follicle at the peak of their
maturity. If the new peritoneoscope can be developed
History of IVF and embryo transfer
along the lines of the operating cystoscope, laparotomy
may even be dispensed with. What a boon for the barren
woman with closed tubes!
Who was this anonymous writer and what is the ‘electri-
cal sign’? John Rock, a gynaecologist at the Free Hospital for
Women, Boston and Harvard Medical School, subsequently
admitted he was the author (Marsh and Ronner, 2008). In
the same issue of New England Journal of Medicine, Rock
described the use of a potentiometer to detect the time
of ovulation (Rock et al., 1937). At this time it was not
known with certainty whether ovulation was associated with
menstruation or whether it occurred mid-cycle between
two menstrual periods. The technique would facilitate the
collection of living human oocytes shortly after their release
from the ovary.
The following year, Rock embarked on two parallel lines
of research. One was the collection and study of the earliest
stages of human development, and the other was to fertilize
human eggs in vitro. He hired Miriam Menkin, who had been
an assistant several years earlier in Pincus’s laboratory at
Harvard, working on the isolation of the two pituitary hor-
mones FSH and LH (McLaughlin, 1982). While there she
learnt to manipulate rabbit ova and embryos. Rock’s work
on early human development continued for about 15 years
in collaboration with Arthur Hertig, a pathologist at Harvard
Medical School, and led to the famous collection of early
human stages frequently referred to in nearly all textbooks
on human embryology (Hertig et al., 1956). Their prodigious
work on the fertilization of human ova led to the claim that
human ova had been fertilized in vitro (Rock and Menkin,
1944; Menkin and Rock, 1948). Nearly 800 human follicular
eggs were obtained from women undergoing surgery and
138 of these eggs were exposed to spermatozoa. During this
time Pincus was consulted. Eventually two ova that had
divided into 2-cell stages were obtained and one 3-cell
stage. The claim was made that fertilization had occurred
in vitro. The claims of Menkin and Rock were widely
accepted for a few years and are frequently cited in more
recent times. Although published during the Second World
War, Rock and Menkin’s first paper attracted sufficient
attention to be covered on the first page of Boston Globe.
The newspaper viewed the work favourably, stating that it
would help towards treating serious problems of infertility.
Some, however, objected to the work. Rock, however, had
antagonists at Harvard and their influence is said to have
subverted any further studies on human embryos (McLaugh-
lin, 1982).
Several years later, in 1954, Landrum Shettles, at
Columbia, claimed to have fertilized a human ovum
in vitro by repeating Menkin and Rock’s protocol (Shettles,
1954). In the archives at Harvard Medical School there is a
letter dated 6 June 1954 from Carl Hartman, then Director
of the Ortho Research Foundation, to John Rock. He
wrote:
I don’t believe you ever got in vitro fertilization . . . Have
a dozen reasons to question your conclusions, chief of
which is the simultaneous and independent discovery
by Chang, Austin and Blandau [Braden?] that ‘raw’
sperms won’t fertilize any egg even in vivo! Sperms must
be ‘capacitated’ (Austin) in the female tract, either in
the uterus or the tube.
121
Furthermore, Hartman encouraged Rock to continue his
work, for he continued:
Now, I want you to go back to the problem and clean it up
and really immortalize yourself. Inject 50,000,000 sperm
into a woman’s uterus. In 2 h take out the sperms and
add to the ovarian egg (but only from a 16–18 mm. folli-
cle, eggs in lesser ones are N.G.). I’m betting heavy odds
on the outcome of this experiment.
Rock never took up Hartman’s suggestion. By 1954, Rock
had ceased working on IVF/ET. Perhaps he was discouraged
by the very low success rate of their attempts to achieve
IVF. He was ahead of his time because, in the next two
decades, important advances in reproductive biology made
success much more likely. Nevertheless, Rock’s enthusiasm
for developing IVF/ET for the treatment of infertility
remained because, 4 years later after a paper read by
Landrum Shettles before the New York Obstetrical Society,
he commented:
The time may be rapidly approaching when the poor
woman whose tubes had been excised, yet who still
wants a baby, will rejoice that Dr Shettles will be able
to extract an ovum from her ovary, probably not by lap-
arotomy, but through an operating telescope (which can
be done – we have done it); then fertilize the egg
in vitro by the husband’s spermatozoa; and finally put
it back in the uterus. Thus will he impregnate the woman
in spite of the fact that she has no tubes. (Shettles, 1958)
There is ample evidence that Rock came under pressure
to discontinue work on early human embryos both locally by
his colleagues at Harvard and by the Catholic Church. Mean-
while he had become associated with Pincus in another ‘hot’
subject – birth control and the development of the oral
contraceptive pill. It seems likely that he could not defend
two contentious issues simultaneously.
The sixth and seventh decades of the 20th century pro-
duced advances in understanding the physiology of fertiliza-
tion and preimplantation development. Before this period,
many investigators were convinced that fertilization
in vitro had been achieved by Pincus in the rabbit and Men-
kin and Rock in the human. The claims were questioned
after it was discovered in 1951 by two independent groups,
Austin (1951) in Sydney, Australia and Chang (1951) in
Worcester, Massachusetts, that freshly ejaculated sperma-
tozoa could not immediately fertilize an egg since they
require a period of so-called maturation in the female gen-
ital tract. The word ‘capacitation’ was coined to denote the
phenomenon by Austin (1952). How then did Pincus and
Enzmann (1934) get young when they added spermatozoa
to the ova immediately after the spermatozoa were col-
lected? One possibility is that spermatozoa were carried into
the uterus at transfer where they capacitated and then fertil-
ized the ovum in vivo. The discovery of capacitation led to
controversy over the nature of convincing evidence sufficient
to claim successful fertilization of mammalian eggs.
Rothschild (1969) defined fertilization as the incitement
of an egg to development by a spermatozoon, together with
the transmission of male hereditary material into the egg.
The difficulty of unequivocally demonstrating that fertiliza-
tion has occurred was emphasized by Austin (1961) who pro-
posed the following requirements: (i) use of capacitated
122
spermatozoa; (ii) avoidance of aged ova; (iii) confirmation
that a spermatozoon had entered the ovum; and (iv) condi-
tions that exclude parthenogenesis. Parthenogenesis cannot
be excluded by observing preimplantation stages of preg-
nancy since parthenogenotes can develop through these
stages and beyond, though parental imprinting deficiencies
precludes their development to term (reviewed by Markert,
1988). Thus, the ultimate requirement is the birth of off-
spring with the use of genetic markers that demonstrate
characteristics transmitted by the spermatozoon.
Many claims that fertilization in vitro had been success-
ful were made by this time, but all can be criticized on the
grounds that the reported evidence was insufficient. Austin
(1961) cites about 30 papers in which it is claimed that fer-
tilization in vitro was successful in the rabbit, guinea-pig,
human and sheep. He concluded that only the work of
Thibault and his colleagues, Moricard (1954) and Chang
(1959), merited serious consideration. For example,
Thibault et al. (1954) used capacitated spermatozoa to
fertilize rabbit ova. They reported only cytological observa-
tions on the cleaving embryos and did not transfer the
embryos into uterine foster mothers to see if they would
develop into newborn young. Moricard also did similar
experiments without confirming that fertilization occurred
by using embryo transfer. It is now generally accepted that
the first unequivocal achievement of IVF was done in the
rabbit by Chang (1959), working at the Worcester
Foundation where Pincus was Director. This does not mean
that others had not previously achieved IVF; it means that
the necessary rigorous scientific proof was not provided.
In all this early work, IVF was done using biological fluids
of unknown composition as the culture medium. IVF, using
a chemically defined culture medium, was achieved in the
mouse by David Whittingham in 1968 using a medium for
embryo culture described by Whitten and Biggers (1968).
In his book The Eggs of Mammals, Pincus (1936a,b)
described in detail methods for the culture of preimplanta-
tion embryos. He used media based on biological fluids, such
as blood serum, employed at the time in the field of tissue
culture. Some successes were reported, including some pio-
neering attempts at determining the nutritional require-
ments of preimplantation embryos in vitro. A major
development occurred in the field of tissue culture when
chemically defined media were developed by White (1946)
at the Jackson Laboratory in Maine and for animal cells by
Fischer (1947) in Germany. These media allowed culture
experiments to be repeated in different laboratories under
relatively comparable conditions, which is not possible with
biological fluids.
A little-known letter was published in 1947 by John Ham-
mond, Jr, who showed that 8-cell mouse embryos would
develop into blastocysts when cultured in Krebs-Ringer
bicarbonate supplemented with egg white (Hammond,
1947). Whitten in 1956 replaced the egg white with bovine
serum albumin to produce the first chemically defined
medium that supported development of 8-cell mouse
embryos into blastocysts (Whitten, 1956). Earlier stages
would not develop under these conditions. In a totally unre-
lated study of the classic nature versus nurture problem,
McLaren and Michie (1956) had optimized the technique
for embryo transfer in mice. McLaren and Biggers (1958)
used this optimized protocol to transfer blastocysts,
JD Biggers
produced from the 8-cell stage in Whitten’s medium, into
uterine foster mothers, and showed using coat colour as a
genetic marker that phenotypically normal mice developed
from the cultured embryos. The scientific report published
in Nature was greeted in the London Daily Telegraph under
the heading ‘Brave new mice’. We received no adverse crit-
icism of the work, although a well-known scientist from
Cambridge, Cecilia Lutwak-Mann chastised us by mail for
allowing our results to be published in the popular press.
We were not interested, like Pincus 20 years earlier, in sen-
sational implications of our work, but only in the scientific
applications. In the popular magazine Discovery we wrote:
It is inevitable that the thoughts of anyone who has
worked on the subjects outlined in this article should
turn to Aldous Huxley’s fantasy ‘Brave New World’,
where he describes completely artificial fertilization
and development of human embryos. Fortunately we
are far removed from this frightening prospect. The
study of the cultivation and transfer of embryos is none
the less of the greatest interest, both from the point of
view of pure science, and because the techniques associ-
ated with it are potentially of immense value in the
investigation of many biological problems in medicine
and agriculture. (Biggers and McLaren, 1958)
This work first demonstrated that live-born mice could
develop after being exposed to chemically defined media
during the initial stages of development. While this work
was ongoing, Whitten (1957) found that 2-cell mouse
embryos would also develop into blastocysts if lactate was
added to the medium. A few years later, Biggers et al.
(1967) showed that the first cleavage division also required
pyruvate in the medium. These observations established the
basis for the design of several media, including medium
KSOM/AA that many use to culture mouse embryos today
and which has been slightly modified in one commercially
available medium for the culture of human embryos
(Global: IVFOnline, Guelph, Ontario, Canada).
Successful IVF requires the availability of ova ready for
fertilization. Pincus and Enzmann (1935) in the rabbit and
Pincus and Saunders (1939) in the human had shown that
oocytes isolated at the germinal vesicle stage would pro-
ceed to the metaphase-II stage spontaneously when placed
in culture. In the next 30 years, the phenomenon was dem-
onstrated in several more species: mouse, rat, hamster,
rabbit, sheep, cow, pig and monkey (reviewed by Biggers,
1972). Particularly important contributions on larger ani-
mals with longer maturation times were made by Edwards
(1962, 1965a), who observed the phenomenon using
serum-supplemented cell-culture media. Another advance
was made by Kennedy and Donahue (1969), who showed
that human oocytes could complete meiosis in several
chemically defined media, including those commercially
available for cell culture. These media varied in complexity,
from F10, which contains many components, to simple
media with relatively few components based on modified,
pyruvate-supplemented Krebs-Ringer bicarbonate.
Pincus (1940a,b), in another pioneer paper using the rab-
bit, described research results with the objectives ‘to ripen
as large a number of follicles as possible and to determine
whether the ova obtained at ovulation were fertilizable’.
The technique was needed to increase numbers of ova and
History of IVF and embryo transfer
early embryos for experimental study. He succeeded in
devising a technique, using crude extracts of the pituitary
gland that satisfied these objectives and thus was the first
to describe the technique of superovulation widely used
today. By this time, it was known that the maturation of
Graafian follicles and the ovum depended on two hormones
from the anterior pituitary gland originally called ‘Prolan A’
and ‘Prolan B’ by Zondek (1929), now known as FSH and LH
(reviewed by Lunenfeld, 2004). An important study by
Fowler and Edwards (1957) worked out the protocol used
to this day for the production of synchronous mouse oocytes
capable of fertilization and development to term, using the
sequential injection of pregnant mare serum gonadotrophin
and human chorionic gonadotrophin. By the time Edwards
began his work on the maturation of human oocytes
in vitro, human menopausal gonadotrophin (a mixture of
FSH and LH) and chorionic gonadotrophin (mainly LH) were
available to stimulate follicular growth (Edwards et al.,
1970).
Another technique that has played an important role in
human IVF is laparoscopy for the recovery of oocytes from
the mature Graafian follicle. The idea of an ‘operating tele-
scope’ originated early in the 20th century (Steptoe, 1967).
Its potential usefulness to recover human oocytes was rec-
ognized by Rock in his editorial in New England Journal of
Medicine in 1937. A cystoscope employed in urology, similar
to a laparoscope, was first used to recover an oocyte from a
single human follicle in France by Klein and Palmer (1961).
Putting it all together: Robert Edwards’
contributions
Interest in human in IVF/ET recurred in the 1960s largely
due to the work of Robert Edwards. In the course of a few
years, Edwards and his colleagues published the following
key papers that paved the way for the birth of the first
‘test-tube’ baby:
(1) ‘Maturation in vitro of human ovarian oocytes’ in The
Lancet (Edwards, 1965b).
(2) ‘Early stages of fertilization in vitro of human oocytes
matured in vitro’ in Nature (Edwards et al., 1969).
(3) ‘Fertilization and cleavage in vitro of preovular human
oocytes’ in Nature (Edwards et al., 1970).
(4) ‘Laparoscopic recovery of preovulatory human
oocytes after priming of ovaries with gonadotrophins’
in The Lancet (Steptoe and Edwards, 1970).
Edwards’ interest in the mechanisms involved in the
establishment of pregnancy arose from cytogenetic studies
on oocyte maturation (reviewed by Edwards, 2001; Johnson,
2011). He extended work done in the 1930s by Pincus and his
colleagues by showing that spontaneous maturation
occurred in several species using the chemically defined
medium 199 supplemented with 15% serum. Edwards began
his initial studies on the maturation of human oocytes by
incubating them for only 12 h, following Pincus and
Saunders (1939). This period of incubation was too short,
and it took some time before Edwards found that the
required time was between 36 and 43 h after laparoscopy.
Obtaining human oocytes for experimentation is difficult.
123
To get access to further material, Edwards spent 6 weeks
at the Women’s Clinic in Johns Hopkins Hospital, Baltimore
with Howard Jones and Georgeana Seeger Jones. This
rewarding visit resulted in key paper 1, which was published
from Johns Hopkins Hospital. The availability of meiotically
mature human ova that resulted from this work opened the
door to needed experimental work on the fertilization of
human eggs in vitro. Unfortunately, attempts to fertilize
these in-vitro matured oocytes with capacitated spermato-
zoa consistently failed (Edwards et al., 1966).
Fertilization in vitro of matured human oocytes was
finally reported in key paper 2. It occurred almost serendip-
itously. Bavister was working on the capacitation of hamster
spermatozoa in a laboratory adjacent to Edwards’ labora-
tory. Capacitation in vitro had been observed in the hamster
by Yanagamachi and Chang (1963) using a simple modified
Tyrode’s balanced salt solution. Bavister’s attempts to
repeat this work resulted in erratic outcomes that he
showed were due to poor pH control. A modified medium
supplemented with pyruvate and bovine serum albumin
was finally produced in which the pH was strictly controlled
at 7.6. When in-vitro matured human oocytes were incu-
bated for 6 hours with human spermatozoa in this medium,
spermatozoa were observed inside the oocytes, and one
oocyte contained two pronuclei. In a letter to Nature,
Rothschild (1969) pointed out that these experiments dem-
onstrated only the initial stages of fertilization and did not
satisfy the stringent requirements of Austin (1961); the
results provided no proof that normal young would finally
develop. Edwards and colleagues then showed that human
ova penetrated by a spermatozoon could develop in modi-
fied Bavister’s medium and other media through the
subsequent cleavage divisions to the blastocyst stage (key
paper 3). Bavister’s original formulation was modified by
increasing slightly the sodium and potassium concentra-
tions. A second, simple, chemically defined medium was
also used; the medium had been described by Whitten and
Biggers (1968) for the complete culture of mouse embryos
from the zygote to the blastocyst thus overcoming the 2-cell
block. This medium was made available before publication
to Whittingham (1968) for his successful studies on IVF in
the mouse, and it was subsequently passed on to Edwards
and his colleagues. Three other media were widely used in
cell culture – Ham’s F10, Eagle’s 199 and Weymouth’s
(modified by the addition of pyruvate) – all supplemented
with inactivated fetal calf serum. Pyruvate was included
in all of these media after the demonstration in the mouse
that it is required for oocyte maturation and the first cleav-
age division (Biggers et al., 1967). Some penetration of
spermatozoa into the ova, pronuclear formation and cleav-
age were observed with all media, although at low rates.
These experiments did not rule out the possibility of parthe-
nogenesis. Thus, it was not until 1978 and the birth of the
first baby produced by IVF/ET that normal IVF in humans
was finally proved.
A cohort of oocytes recovered from an ovary are invari-
ably at different stages of maturation. Key paper 4
describes the use of two gonadotrophins given sequentially
to synchronize as closely as possible a cohort of oocytes
at a stage normally reached just before the expected time
of ovulation. The two hormones were human menopausal
gonadotrophin (HMG) and human chorionic gonadotrophin
124
(HCG). The preovulatory oocytes were then harvested for
IVF/ET by puncturing the follicles with a laparoscope and
aspirating them with a specially designed piece of
equipment.
A period of about 8 years elapsed before the first baby
conceived by IVF was born in July 1978 (Steptoe and
Edwards, 1978). In all but one case, the patients failed to
become pregnant after embryos produced by IVF were
transferred to the mother. The one exception resulted in
an ectopic pregnancy. Various reasons for the failure of
transferred cleavage-stage embryos to develop were consid-
ered. For example, such transfers in mice to the uterus
rather than to the oviduct were rarely successful due to
the inappropriate hormonal priming. However, work on
the transfer of cleaving human embryos to the uterus was
encouraged by the report of Marston et al. (1977) who
showed that a 5-cell rhesus monkey embryo could develop
into a newborn baby after transfer into the uterus of its
mother. Another potential contributory cause of the failures
in human patients was the trauma involved in transferring
the embryos into the uterus. A further possible cause was
the endocrine disturbances elicited by the HMG and HCG
used for ovarian stimulation, leading to what Edwards called
luteal weakness (Edwards, 2001). After trying various hor-
monal supplements, Edwards and Steptoe turned to the
recovery of a single oocyte from a natural cycle by monitor-
ing the patient’s LH surge near the time of ovulation. The
first IVF baby, Louise Brown, was conceived in this way.
The first babies produced following ovarian stimulation
using clomiphene and HCG were not reported until 1981
by Carl Wood’s group (Trounson et al., 1981).
Edwards’ papers generated renewed interest in IVF in the
USA, particularly that of Howard Jones and Georgeanna See-
ger Jones at the Woman’s Clinic at Johns Hopkins Hospital.
Others who became interested in the field were Pierre Sou-
part at Vanderbilt School of Medicine, Nashville (Soupart
and Morgenstern, 1973; Soupart and Strong, 1974) and Mel-
vin Taymor at the Brigham Hospital in Boston (Berger et al.,
1975). Soupart, in fact, had a grant application approved by
a study section at the National Institutes of Health to sup-
port human IVF, an application which, in an unusual step,
had been referred to an Ethics Panel. Unfortunately Soupart
died before approval was given.
By the end of the 1960s, Edwards decided to seek
large-scale support for his work, and in 1971 he and Steptoe
applied to the British Medical Research Council for a grant
to set up a clinical research programme, but was rejected,
a great disappointment to Edwards. In retrospect it may seem
that those who decided the fate of the application were short
sighted. Recently, Martin Johnson of the University of Cam-
bridge and colleagues reviewed the deliberations within the
Medical Research Council (Johnson et al., 2010). Their
research revealed a complex network of conflicting interests,
mainly administrative but not scientific, that resulted in the
rejection. On the whole, the scientific advisors to the MRC
approved of the science, although concern was expressed
over the potential of producing birth defects. One reviewer
suggested the technique should first be evaluated in mon-
keys. Rejection was due to an excessive budget, concern over
what was perceived as the use of patients in clinical research,
doubts about the high media profile of Edwards and Steptoe,
with some hints of politicking by aspirant Directors of other
JD Biggers
MRC-supported organizations who vied for the clinical
research programme in their own establishment. Edwards
may well have unwittingly killed his application by insisting
that if the clinic was located at a site other than Cambridge
he was not interested. In 1979, after the birth of Louise
Brown, Edwards applied to the National Health System for
state support to establish an IVF clinic, but again the applica-
tion was rejected. He finally secured venture capital to set up
Bourn Hall, the first IVF clinic, which was opened in Septem-
ber 1980 located on an estate near Cambridge. Clinical
research proceeded rapidly in this new environment and it
included the development of successful treatment regimens
for overcoming the luteal weakness that plagued the early
work. Securing government support in this field remained dif-
ficult. A second application for research funding on IVF to the
Medical Research Council also failed. Nevertheless, over
1000 babies were born using IVF/ET in the first 10 years of
Bourn Hall’s existence (Edwards, 2001).
A controversial claim was made in India, soon after the
birth of Louise Brown, that a baby girl had been born follow-
ing IVF/ET. The team that made this claim were two physi-
cians (Subhas Mukerji, leader of the team, and SK
Bhattacharya) and a cryobiologist (Sunit Mukherjee). Mukerji
had done research on hormone assaying at the University of
Edinburgh, and Mukherjee was well trained in cryogenics at
Cornell University. The case was not described in profes-
sional journals but was widely reported with considerable
hoopla in the Indian press (Kumar, 1997). A six-page detailed
report appeared in the popular Indian magazine Sunday
under the title ‘Our very own test-tube baby’ (Mitra and
McMahon, 1978). The baby was born on 3 October 1978 and
thus, if the claim is true, her mother became pregnant about
6 months before Louise Brown was born. The case also
attracted considerable attention since, for the first time,
ovarian stimulation and cryopreservation of the embryo
were reported. An official at the NIH asked me to look into
the claim when I visited India to attend the International
Congress of Hormonal Steroids held in New Delhi in the late
fall of 1978. Dr Kenneth Ryan, Harvard Medical School, Dr
Liselotte Mettler, University of Kiel, both also attending
the Conference, and I interviewed Subhas Mukerji in the
Ashoka Hotel on 2 November 1978. He gave us full details
of the protocol that was followed. We were informed that
we could not see the baby since her parents wished to remain
anonymous. The protocol used seemed to follow the prac-
tices used at the time so we were not inclined to dismiss
the claim out of hand. Nevertheless, the Indian doctors we
talked to unanimously regarded the claim as fraudulent. Dur-
ing the next 20 years, the case was largely forgotten and
considered questionable. Then, in 1997, Anand Kumar, a
prominent Indian gynaecologist (Metha, 2010), re-examined
the case in minute detail. He had gained access to the
group’s laboratory and clinical notes and official government
documents. He concluded that Mukerji and his team had
indeed succeeded in producing a baby by IVF/ET and that
bureaucratic interference had prevented Mukerji from justi-
fying his claim in front of his scientific peers. Mukerji had
summarized the details of the case in a letter dated 1
December 1978 to the Director of Health Services of the
West Bengal Government, and on 28 December 1978 the
Director specifically ordered Mukerji not to attend any
conference without prior permission. Permission to accept
History of IVF and embryo transfer
an invitation to present his case in Japan was denied (Anand
Kumar, 1997). Mukerji’s death soon afterwards deprived him
of any further opportunity to justify his claim.
Ethical and moral issues raised in the USA and
the formation of the first clinic in Norfolk,
Virginia
The birth of Louise Brown was widely reported in the US
press, often in a sensational manner. The negative recep-
tion received by John Rock 34 years earlier was repeated
with greater intensity. Based on my own experiences,
I can provide some understanding of the environment that
scientists and physicians encountered in the USA.
In 1978, I was director of a programme project at Har-
vard, supported by the Institute of Child Health and Human
Development, on the biology of early pregnancy. One section
of the project, headed by Dr Melvin Taymor of the Brigham
and Women’s Hospital, was concerned with trying to mature
human oocytes in vitro. The day the birth of Louise Brown
was announced, I received a call from an NIH official freezing
these funds. It was clear that some members of Congress
were upset by the demonstration that a new individual could
be created in a test tube. Soon after, Joseph Califano, Sec-
retary of the then Department of Health, Education and
Welfare, activated a dormant Ethics Board to advise him
and President Carter on whether it was ethically acceptable
for the Federal Government to support work on IVF/ET. I was
appointed scientific advisor to the Board, together with a
philosopher and two lawyers. The members of the Board
were a very talented group of people representative of many
walks of life. Several hearings open to the public were held
by the Board, which eventually recommended that the Fed-
eral Government could support work on IVF/ET subjected to
certain restrictions. In retrospect, it is interesting that one
provision made by the Board was that in any experiment
the spermatozoon and egg must be donated by a married
couple. The recommendations of the Board were never
accepted by the Government, however, and as far as I know
the report still sits in some government pigeon hole (Biggers,
1978, 1981). In 1980, the National Institute of Child Health
and Human Development invited me and Luigi Mastroianni,
Chairman of Obstetrics and Gynecology at the University of
Pennsylvania, to organize a small conference on the
Bethesda Campus about IVF/ET as practised in animals, with
the explicit instructions that discussion of human IVF/ET be
disallowed (Mastroianni and Biggers, 1981). I was chairing a
session when Barry Bavister jumped up at a point in the dis-
cussion and said ‘Let’s talk about the human’. After receiv-
ing frantic signals from the NIH staff in the audience, I
adjourned the meeting for lunch, with the result that my
friends accused me of stifling free speech. When the pro-
ceedings were published by Plenum Press, we were required
not to acknowledge that the NICHD sponsored the meeting.
Clearly the NICHD staff was scared by the possible reactions
of some politicians in Congress.
Soon after the birth of Louise Brown, the President of the
Eastern Virginia School of Medicine invited Howard Jones
and Georgeanna Seeger Jones to come out of retirement
and rekindle their interest in IVF to form an IVF/ET clinic.
A legal requirement for setting up a new clinic was getting
125
a Certificate of Need from the State of Virginia. This
involved having a public hearing. Howard Jones invited me
to testify at this hearing. It turned out to be a horrendous
experience. The hearing took place in a hall that held
200–300 people. The audience had seated themselves in
two groups on opposite sides of the hall, one group support-
ing IVF/ET, including couples who had not been able to con-
ceive, and the other group, largely right-to-lifers, strongly
opposing IVF/ET. The entire meeting turned out to be a
shouting match, the two groups hurling insults at each
other. I particularly remember a right-to-life activist from
Chicago whose purpose was to attack in a particularly insult-
ing way the motives of Georgeanna Seeger Jones, one of the
most dedicated clinicians you could ever meet. Fortunately,
the Certificate of Need was granted. Attacks on the pro-
posed clinic continued to be made in the local media and
this resulted in a lawsuit, which the Medical School won.
The monetary settlement helped create the first IVF clinic
in the USA. Soon after the clinic was formed, the Virginia
Bar Association sponsored a 1-day symposium on IVF/ET.
I was invited to present the first paper outlining the princi-
ples of IVF to the audience of lawyers. The meeting was held
at Virginia Beach, which is also the home of the Christian
Broadcasting Network run by Pat Robertson. His network
organized a protest outside the hotel where the Conference
was to take place. As a result, all speakers were taken to the
back of the hotel and admitted through the kitchen.
Public policy makers like to make the distinction
between a procedure done for the benefit to the patient
and a procedure done for research. In some situations
including IVF/ET, the former is legal while the latter is not.
I remember participating in a long, tedious conference call
with Dr Taymor, who succeeded in producing the first test
tube baby in Boston, and the District Attorney of Boston
over how a particular procedure should be classified. No
clear rules developed other than all issues should be
decided on a case-by-case basis.
Conclusion
IVF and embryo transfer in the human is built upon exten-
sive basic research done by many investigators in reproduc-
tive biology for over a century. Robert Edwards built on this
earlier work, making major scientific contributions. Equally
important was his dogged resistance to those who opposed
IVF/ET on ethical and moral grounds. His unwavering enthu-
siasm and perseverance led to a revolution in the treatment
of human infertility and the establishment of a new branch
of medicine. John Rock was a visionary who was ahead of his
time because the necessary basic science in the field had
not been done. An organization called IVF-Worldwide
attempts to accumulate statistics on IVF around the world.
At the latest count, there are at least 3221 clinics located in
almost all countries in the world and almost 4 million IVF
babies have been born.
Presentation
This article is based on a plenary lecture given to the Soci-
ety for the Study of Reproduction on 1 August 2011, in Port-
land, Oregon.
126
Acknowledgements
The author is indebted to Drs Carol Kountz and Betsey Wil-
liams for help in researching and preparing this paper.
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Declaration: The author is a consultant to IVFOnline, Guelph,
Ontario, Canada.
Received 9 January 2012; refereed 24 April 2012; accepted 24 April
2012.