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Frozen embryo twins born from frozen stored semen

The first twins born from frozen sperm and from frozen embryos in Britain were revealed in January 2001. Stephen Mercer, the father, was infertile after earlier cancer treatment, having been diagnosed with a lymph cancer in 1987 at the age of nineteen. As a precaution, and prior to treatment, he had had his sperm stored in a liquid nitrogen system. This sperm was retrieved some eleven years later, and was then thawed and used to fertilise his wife’s eggs via intracytoplasmic sperm-injection (ICSI) as part on an IVF programme.

Because his wife, Helen Mercer, had an over reaction to fertility drugs, the team from St Mary’s Hospital in Manchester, UK, felt that waiting some months would improve the chances of successful implantation.

Accordingly the embryo blastocysts were frozen in a Planer controlled rate freezer and stored cryogenically for four months before their successful implantation. Greg Horne, senior embryologist, felt that it would have been unwise to implant the embryos at once so they had to be frozen. The twins Abigail and Gabrielle are now eighteen months old.

First Frozen Bone Marrow Transplant in Romania

Autologous bone marrow transplants for patients undergoing chemotherapy have been growing in importance in Europe, Japan and the Far East. However developing countries have often lacked the resources necessary for this kind of intervention.

It is pleasing therefore that Mr Nicolae Tarcea reports that the first such transplant was made in Romania on 5th of June 2001, by a team from the Paediatrics Clinic in Timisoara. The operation was carried out by Prof. Margit Serban and Mr. Virgil Paunescu and the medical centre there used a Planer programmable rate freezer with a 16 litre capacity which had been installed in September 2000.

The transplant was made in collaboration with a team of doctors from the Medullar Transplant Centre from Miskolc in Hungary, a nearby centre with good experience in this discipline and the team was supervised by Prof. Nagy Kalman.

The transplant was made for a twelve year old child diagnosed with Hodgkin’s lymphoma, who had two previous relapses on record. The operation appears to have been successful, the first signs of grafting appearing after ten days post transplant. The child is said to be feeling well, and the indications are hopeful.

Frozen Ovarian Tissue success for British medical equipment

Frozen Ovarian Tissue success for British medical equipmentThe recent birth in Belgium of a healthy baby girl, Tamara, has caused excitement throughout the world. Tamara is the first baby to be born to a woman after a frozen ovarian tissue transplant and despite the mother, Ms Touirat, being in a state of premature menopause provoked by the chemotherapy and the radiotherapy that had cured her of the cancer.

The revolutionary treatment was carried out by Professor Jacques Donnez and his team at the Cliniques Universitaires Saint-Luc with help from a new and sophisticated freezing equipment made by a British company. By the year 2010 it is estimated that about one in 250 people in the adult population will be childhood cancer survivors. Whilst 90% success rates can prevail for radio and chemotherapy treatment in such childhood treatments, ovaries are very sensitive to cytotoxic treatment and generally fertility function is compromised. The restoration of the ability to have children naturally after treatment is a high medical priority.

In this ground breaking case, part of Ms Touirat's left ovary was removed and the tissue was controlled rate frozen in a Planer step freezer before being stored in liquid nitrogen whilst therapy was undertaken. Some six years later tissue was implanted near Ms Touirat's fallopian tube and eggs were shortly produced, allowing a normal conception then to take place; on September 23, 2004 Tamara was finally born, weighing 3.72 kg. In their paper in the scientific journal The Lancet, the researchers say all the evidence they can see shows that the egg follicle, which ripened into the egg during the menstrual cycle in which Mrs Touirat became pregnant, came from the transplanted tissue.

The deep freezing of biological samples in liquid nitrogen has been used in hospital and veterinary medicine for some while. However, not all biological samples accept this process easily and Planer plc, a UK company based near London’s Heathrow airport, has been pioneering the use of specialist equipment for controlled rate freezing for these difficult but vital samples over the last thirty years. Its programmable step rate freezers allow the viable cryogenic preservation of samples such as skin, blood vessels, bone marrow, cord blood and other tissue that would otherwise not survive for long term storage in liquid nitrogen.

While sperm and embryos freeze and thaw relatively well, repeatability in human oocyte freezing is harder. However dramatic progress in oocyte preservation was also announced in September 2004 by Italian researchers Dr Raffaella Fabbri and her team. They say that they have achieved a breakthrough of thirteen live births using eggs that had been frozen from 68 different women. They also used the new versatile Planer freezer, similar to the one used by Prof Donnez. Their viability rate of 37% is a big step forward, but Dr Giovanni Coticchio leader of the research team at Bologna University, said that in future they hope to achieve a 75 to 85 per cent survival rate.

Planer plc was formed in 1973 makes the only freezers that can cope with the differing requirements for embryo, oocytes and tissue freezing. Amongst many other achievements a Planer freezer was used in the first human birth from a frozen embryo in 1983 and also the first human birth from a frozen oocyte in 1997. Thousands of researchers, IVF labs, vets and doctors now use Planer freezers around the world.

Photo shows Ouarda Touirat and Tamara with cryogenic freezing equipment used to freeze her tissue.

Child born after 21 year semen storage using Planer controlled rate freezer

Child born after 21 year semen storage using Planer controlled rate freezer Researchers from St Mary's Hospital and Christie Hospital in Manchester, involved in the treatment of the couple who conceived using sperm frozen for twenty one years , published their case study in the June issue of the journal ‘Human Reproduction’. The baby boy was born after four cycles of IVF, using sperm frozen before his father, now aged 42, underwent treatment for testicular cancer at the age of seventeen.

The couple subsequently embarked upon four cycles of Intracytoplasmic Sperm Injection, better known by its short form ‘ICSI’, a fertility treatment commonly employed where the sperm are too weak to penetrate the egg. In this case although fertilisation and subsequent fresh embryo transfer were achieved in three out of the four cycles no pregnancies were achieved. Success was only finally achieved with the use of the last remaining viable embryo which had been cryopreserved in a Planer controlled rate freezer and placed in a long term liquid nitrogen storage vessel.

Based near London’s Heathrow airport, Planer were pioneering the controlled rate freezer around the same time as the patient’s sperm was originally being frozen. Cryopreservation had its roots in the ‘deep freeezing’ of avian sperm, which was commenced after the Second World War; human pregnancies using frozen sperm followed in 1955. The utilisation of 'rapid immersion in liquid nitrogen' methods for storing delicate and valuable samples did not always produce the necessary viability however and a 'step cooling' process, allowing a ‘difficult to freeze’ sample to equilibrate osmotically in cryoprotectant before cooling over predetermined ramps proved necessary.

Equipment for this controlled rate freezing process proved a spectacular breakthrough in the preservation of biological samples, such as human embryos, and Planer received the Queen's Award for Technology for it in 1984. Controlled rate freezing is now a universally established process for the pre freezing prior to long term storage of specimens ranging from human, such as bone marrow - to botanical, such as special seeds; Planer now sell their specialist freezing machines all over the world.

Scottish Eagle born from frozen sperm

Scottish Eagle born from frozen spermScottish scientists have had a small breakthrough in their efforts to save endangered birds after they successfully bred a golden eagle using frozen sperm. A team from the University of Abertay Dundee managed to engineer the birth of a chick, called Crystal, who was hatched at a breeding centre in South Lanarkshire on 28 April 2004. Dr Graham Wishart, a biologist, and Andrew Knowles-Brown, an eagle breeder inseminated Crystal’s mother using thawed sperm which had been frozen for one month. There are many obstacles to breeding raptors in captivity naturally - the birds may not be ready to breed at the same time and, in the case of endangered species, it is often difficult to match up a breeding pair if the few remaining individuals are scattered across continents.

The freezing of sperm allows storage in near perpetuity, so for rare birds the optimum breeding conditions and any trans national programme can be managed. Before breeding Crystal, the Dundee team worked to improve the technique it used to breed an eagle using cryo-preserved sperm in May 2001, a world first. That eagle, a cross between a golden and a steppe eagle, was bred using sperm frozen at a slow rate. Following talks with raptor breeding experts in Spain, Dr Wishart and Mr Knowles-Brown used a Planer Kryo 10 controlled rate freezer to experiment with different cryo preservatives and freezing rates to obtain the optimal protocol for the process which was then applied via field freezing at the aviary concerned.

Homograft Valve Bank: Our Experience in Valve Banking

Author(s):Susan Verghese, P Padmaja, B Sindhu, SJ Elizabeth, Nancy Lesley, KM Cherian

Homograft Valve Bank: Our Experience in Valve BankingBackground: A cardiac homograft valve bank with cryopreservation facility was established at the Institute of Cardiovascular Diseases, Madras Medical Mission, Chennai in July 1995. Methods and Results: During the last 7½ years of its existence, from July 1995 to March 2003, 588 hearts were processed. The valves harvested were 390 aortic, 400 pulmonary and 39 others including mitral valve, aortic conduits, pericardium etc.; 176 (29.9%) hearts were discarded for various reasons which included failure to sterilize, HBV, HIV, HCV, treponema pallidum hemagglutination test positivity, atheromatous/fatty streaks, incompetent valves, and dissection mistakes. The valves were sterilized using an antibiotic cocktail of vancomycin, amikacin, streptomycin, cefotaxime and amphotericin B in Hank’s balanced salt solution.

Of the 585 valves issued for clinical use, 247 were aortic, 323 pulmonary and 15 others (mitral valve, pericardium, conduits). Gram negative bacilli were the predominant contaminants from the hearts during the first half (July 1995 to February 1999) and gram positive organisms were the predominant contaminants during the later half (March 1999 to March 2003) of the study period. A variety of fungal contaminants like candida, aspergillus, penicillium and other fungi were also isolated from the homograft hearts at procurement. The valves were used most commonly for Rastelli procedure/right ventricular-pulmonary artery conduit (48.71%) followed by Ross procedure (23.41%).

The other procedures were aortic valve replacement (6.15%), truncus repair (5.81%), unifocalization with conduit repair (6.49%), aortoplasty (0.512%), left ventricular-pulmonary artery conduit (0.512%), pulmonary valve replacement (0.512%), aneurysm repair (0.34%), Norwood repair (0.34%), mitral valve replacement (0.17%) and other procedures (7%).

Department of Cardiology, International Center for Cardiothoracic and Vascular Diseases and Madras Medical Mission, Chennai.

Stem Cells: All India Institute of Medical Sciences

The All India Institute of Medical Sciences, New Delhi - (AIIMS) - recently announced its pioneering stem cell treatment using an injection method whereby stem cells are placed directly into the patient. In a ground breaking study, under its Director Dr. P. Venugopal, over two years up to January 2005, thirty five cardiac patients were given stem cell treatments and monitored regularly. There have been real improvements and no mortalities, despite the fact that all the patients were brought in at a stage when their hearts were deemed beyond bypass surgery. A national stem cell centre will now be started at AIIMS which will co-ordinate the research and its applications.

The Institute, a world-wide centre of excellence, is a user of several Planer machines which were supplied by Indus Instruments Impex P Ltd. Stem cell freezing is as yet only planned but Planer Kryo 560s and 360s are already used in homograft, HIV programmes research and bio research. Mr. P. P. Venkitachalam, Managing Director of Indus, said that "We very much hope that also in the pipeline is an umbilical cord cell bank". This would be a part of the stem-cell facility at AIIMS.

In cord blood banking, blood is retrieved from umbilical cords of neonates which is conventionally discarded along with the placenta. The cells are then treated and controlled rate frozen prior to long term cryo-preservation in liquid nitrogen These, on purification, are a rich source of stem cells. Disorders which can be treated with stem cells from umbilical cord blood include various genetic disorders that affect the blood and the immune system, leukaemia and certain cancers as well as some inherited disorders. Cord blood stem cells can benefit a wide range of recipients including the donor, his siblings, other family members and even unrelated recipients.

Second frozen ovarian tissue transplant

Second frozen ovarian tissue transplantA team from the University of Louvain in Brussels announced that a second woman in their care has had a successful ovarian tissue transplant. Announcing the details of the case at the 12th World Congress on Human Reproduction in Venice 2005, they said the woman's menstrual cycle, which had stopped after radiotherapy treatment, had been restored. The principle of this type of procedure is to re-implant ovarian tissue into the pelvic cavity (orthotopic site) or to a heterotopic site, like the forearm, once treatment is completed and the patient is disease-free.

In 2004, in a pioneering procedure, the same team, led by Professor Donnez, treated Ourda Touirat, a thirty two year old cancer patient. She had a daughter in September 2004 after undergoing the transplant. At the time some experts suggested that it was possible her menstrual cycle might have restarted naturally - but this latest success would indicate that this was not the case. The latest, as yet unnamed, patient is a twenty eight year old woman who had some of her ovarian tissue removed before receiving radiotherapy treatment for sickle-cell anaemia; this treatment would normally make a woman infertile. In the transplant procedure tiny strips of ovarian tissue, one to two millimetres thick, were taken from the most productive part of the ovary. These were then cut into sections and frozen in a Planer controlled rate freezer to liquid nitrogen temperatures.

The patient's tissue was removed in 1999, before her radiotherapy treatment and was stored in liquid nitrogen after being controlled rate frozen. This is similar to the ‘banking’ of autologous bone marrow, also using Planer freezers, which is carried out prior to irradiation in some cancer treatments. In August 2004 some of the tissue was re-implanted next to one of her ovaries and she began menstruating again in January 2005. Professor Jacques Donnez, who has three Planer freezers, told the Human Fertility and Embryology conference: "We are hoping that she will become pregnant like the last patient, but we do not know how long that may take. This is just the beginning of the story, but it does prove that for a second woman we have been able to restore ovarian function." He added that the patient was delighted with the success of the procedure so far.

It is estimated that by the year 2010 up to one in 250 adults will be the survivors of childhood cancers. The potential options available to preserve fertility in female patients facing premature ovarian failure include immature and mature oocyte cryopreservation, and embryo cryopreservation. For patients who need immediate chemotherapy cryopreservation, of ovarian tissue is a possible alternative. So far 146 other patients of Professor Donnez have had ovarian tissue cryopreserved before starting chemotherapy.

(The photograph shows the Louvain cryopreservation lab, under Dr Demylle, with two Planer Controlled Rate Freezers.)

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