Tap chi Cong nghi Sinh hgc 10(4): 625-632, 2012
REVIEW
T H E 2 1 ^ C E N T U R Y A S P E C T O F E M B R Y O A N D S T E M C E L L T E C H N O L O G I E S
Bui Xuan Nguyen, Nguyen Thi Uoc, Bui Linh Chi, Nguyen Trung Thanh, Nguyen Viet Linh, Dang Nguyen Quang Thanh, Nguyen Van Hanh, Nguyen Thi Men, Tran Thi Thom, Nguyen Thi Hiep, Nguyen Thi Nhung, Nguyen Thi Hong
Insthule of Biotechnology. Viemam Academy of Science and Technology
SUMMARY
The 21st Century is the era of modem biology, of which the development of embryo and stem cell research has a great impact on human being and activities. The 2012 Nobel Prize in Physiology or Medicine was awarded to scientists, who working on cloning and stem cells research, predicting Us burst development in both innovative and frontier aspects in this paper we review previous scientific ideas and discovenes, including the reversible differentiation theory of Spemann in 1932 the set up of human in viiro fertilization (IVF) by Edwards and Steptoe in 1978, the use of nuclear replacement which allowing successful somatic cloning in 1997, the discovery of stem cells in bone minow in 1963 and the induced plunpotent stem cells in 2006. The development of related biotechnologies and their impacis on animal breeding medicine and human health, as well as biodiversity conservation for durable development has been discussed We also present herein (i): the main results of research on embryo transfer embryo production b> IVF and somatic cell nuclear transfer, embryo cryopreservation and sex determination in domestic ind wild animal species, as well as research on human stem cell banking and differentiation which were carried out at the Vietnam Academy of Science and Technology, and (n): iheir potential and challenge in a new local and mttmation i! context Keywords: Embryo. .• ".ells, biotechnology, 2P'
INTRODUCTION
The 2012 Nobel Prize in Physiology or Medicine was awarded lo John B Gurdon, a developmental biologist at the Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Umted Kingdom, and Shinya Yamanaka, a slem cell researcher at the Kyolo University, Japan and the Gladstone Institute, University of California, San Francisco
John B. Gurdon has been considered as one of the pioneers in cloning research. In a classic experiment in 1962, he obtained a normal developed tadpole after replacing the immature cell nucleus of a frog egg wilh the nucleus from a mature intestinal cell and discovered that the specialization of cells is reversible (Gurdon era/,, 1962).
In 2006, S. Yamanaka obtained induced pluripotent stem cells (iPSCs) exhibiting morphology and growth properties indistinguishable from embryonic stem cells (ES) by genetically 'reprogramming' adult somatic cells (fibroblasts) without using oocyte for nuclear iransfer (Takahashi, Yamanaka, 2006).
The mentioned studies of Gurdon and Yamanaka was concerned the embryo and slem cells research, ihe field of which the Nobel Prize in Physiology or Medicine m 1935, 2007, 2009 and 2010 were awarded to Hans Spemann "for his discovery o f t h e organizer effect in embryonic developmeni", to Mario R Capecchi, Martin J Evans and Oliver Smithies "for their discoveries of principles for introducing specific gene modifications in mice by the use of embryonic slem cells", to Elizabeth H.
Blackburn, Carol W Greider and Jack W Szostak
"for the discovery of how chromosomes are protected by telomeres and the enzyme telomerase", and to Robert G Edwards "for the developmeni of in vitro fertilization"
In lis award announcement in 2012, the Nobel committee said that the two scientists' work has
"revolutionized our understanding of how cells and organisms develop". These groundbreaking discoveries open the new horizon for embryo and stem cells technology, predicting the burst development in both innovative and frontier aspects m the twenty-first ceniury.
Research during recent years has shown that 625
Bui Xuan Nguyen et ai human iPSCs could be differentiated to any celt type
in the human body, including sperms and oocytes which capable to be fertilized and developed to new organism We will witness the remarkable progress m many areas of artificial reproduction for infertility treatment, regeneration medicine for patients suffering from degenerative diseases such as Alzheimer's, Parkinson's, diabetes, traumatic spinal cord injLuy, heart disease, hearing and vision loss, and other genetic defect diseases.
In this paper, we review previous scientific ideas and technique progress in embryo and stem cells technologies, impacts and perspectives of their research and application.
Scientific ideas a n d technical progress Embryo and stem cell technologies associate with two important procedures: (i): the use of embryo and stem cells as the first-hfe materials with self-renewal capacity and differentiation and (ii): the transfer or transplantation for replacement, regeneration or restoration.
The world's first successful animal transplantation experiment from fertilized embryos was conducted by Walter Heape, the pioneer in reproductive biology, (Heape, 1891). The first successflil human embryo transfer was performed by Steptoe and Edward nearly 100 years later, in 1978 (Steptoe PC, Edwards, 1978).
Three important scientific ideas-discoveries postulated during the period from 1890 to 1978 are in vitro fertilization (IVF) technique, the use of nuclear replacement for cloning and the detection of stem cells in bone marrow.
The idea ofthe fertilization procedure outside of the body was published in 1932 m the science fiction novel 'Brave New World' written by Aldous Huxley in which, the in vitro developed embryo was cultured to develop completely in glass vessels (Huxley,
1932). In the following years, Pincus (1935) at Harvard University, Charles Thibault, INRA French (1954) and Chang at Boston University (1959) cultured the fertilized eggs of the black rabbit outside the body and later, they successfiilly transplanted them into other white one. These groundbreaking works paved the way for human IVF, which developed by Edwards at the Cambridge University. At early of 1950s, Edwards realized that in vitro fertilization could represent a possibility for infertility treatment and in 1969 he obtained the
human egg fertilized in vitro in a test tube. His efforts were finally crowned by success only 28 years later, when Louis Brown, the world's first "test tube baby" was bom in 1978.
The idea of cloning by nuclear transfer was first reported by Spemann, a German embryologist, who received the Nobel Prize in Medicine in 1935 for the discovery of the organizer effect during embryo development. Disproving the August Weismann's theory that cells lose genetic information with each division, Spemann proposed, in a paper entitled
"Embryonic Development and Induction" published in 1938, the "fantastical experiment" of cloning an organism from differentiated or even from adult cells using nuclear transfer method. However, like Aldous Huxley case, at this time, he did not have facilitated technique to realize his ideas. His proposed cloning procedures have been realized only late in 1952 when Briggs and King from the Institute for Cancer Research, Philadelphia succeeded in getting normal development inlo tadpoles by transplanting a nucleus from a frog blastula cell into an enucleated frog egg (Briggs and King, 1952), and in 1962, when Gurdon obtained the same result, but by transplanting a nucleus from a mature intestine specialized cell derived.
During that time, stem cells from bone marrow were isolated by James Till and Ernest McCuUoch from the Ontario Cancer Institute in Toronto. Working with mice, which received a high dose of bone marrow cells transplantation after treatment with lethal doses of radiation, James Till and Ernest McCulloch observed in the spleens of survival animals the nodules developed from injected cells (Till and McCulloch, 1961). They reported later, in Nature in 1963 that the cells from these nodules were produced by a single stem cell and conserved their self-renewal properties and can differentiate into more specialized cells. Later, in 1981, the isolation and establishment of murine embryonic stem cell lines was independently reported by Martin Evans and Matthew Kaufinan at the University of Cambridge (Evans & Kaufman, 1981) and Gail Martin at the University of California (Gail Martm, 1981).
The establishment of modem research techniques and instruments such as new generation of microscopy associated with micromanipulation systems, confocal microscopy, immunocytogenetic technique, PCR and other DNA techniques during the last decades made the intended improvement in
626
Tgp chi Cdng ngh0 Sinh hgc 10(4). 625-632, 2012 embryo and stem cells technologies, particularly the technique of cloning by nuclear transfer.
Two important events of this period were the birth of Dolly - the first cloned mammal from somatic cells in 1997 (Wilmut et ai. 1997) and the discovery of induced pluripotent stem cells without nuclear transfer by Yanamaka in 2006.
After Dolly, somatic cell nuclear transfer (SCNT) was successfully performed m many other mammal species including mouse, rabbit, cattle, horse bulls, cat, dog, goat, pig, etc. Since SCNT offers the possibility to develop therapeutic cloning for degenerative diseases treatment in humans, the successful somatic cloning in mammalian species marked the beginning of a new era for stem cells technology. By using the patient's own somatic cells as nucleus donor, SCNT allows producing embryonic stem cells, which eliminates the risk of patient immunological immuno-rejection. Production of embryonic stem cells has become research goals in many laboratories and research centers. One year after the bom of Dolly, successful production of human stem cells from IVF embryos was reported by James Thomson from the University of Wisconsin- Madison (Thomson el ai. 1998) However, up to present, the production of stem cells from SCNT embryos resulted only in some animal species, including cattle, pig, mouse and monkey.
SCNT created a new model for further investigation of the underiying molecular mechanisms of pluripotency and self-renewal regulation, leading to a better understanding in the role of oocyte factors involved into the reprogramming of transferred nucleus. Using real- time microscopy and fluorescently tagged histones to study the transcription of mouse nucleus transferred into a frog oocyte, Gurdon (Nobel prize in 2012) and his team recently reported direct evidence of the participation of histone H3,3 deposited by the histone-interact ing protein HIRA as the key factor in reverting nuclei to a pluripotent type (Jullien el ai, 2012).
These results of reprogramming investigations underline the conditions for new approaches of stem ceils production- the induction of pluripotency by introducing of transcription factors. Yamanaka (Nobel Prize in 2012) and his co-workers are the first group successfully created induced pluripotent stem cells (iPS cells) in mice in 2006 and in human in 2007. The information about factors that induce the
reprogramming was limited at this time and Yamanaka used different genes combinations to find finally a combination of foiu" factors, Oct3/4, Sox2, c-Myc,and KIf4 that could reprogram the fibroblasts into immature stem cells. As the induced cells exhibited the properties of ES cells, resulting in tumors with all three germ layers after subcutaneous transplantation into nude mice, their discovery was immediately considered a major breakthrough in which iPSCs might effectively replace embryonic stem cells (ESCs), with the additional advantage of permitting autologous transplant.
Recently, Ring and coworkers used a single gene Sox2 for direct reprogramming of mouse and human fibroblasts into multipotent neural stem cells (iNSCs) capable of soon maturing into neurons and developed into neural networks within a month (Ring era/., 2012).
The integration of frontier and innovative technology: Results and perspectives
Dr. Ahmed Zewail, Nobel Laureate in 1999, in his lecture on science and technology in the 21^' century, gave a new outlook on technologies with three categories: I) the 'simple' technology solving domestic problems of everyday life, 2) the 'innovative' technology which can make participation in the world market possible; and 3) the 'frontier' technology which is concemed with research into the unknown, representing an investment in ihe future.
The research and development of embryo and stem cells during the last decades represents the intensive integration of frontier and innovative technologies.
The IVF technology had developed rapidly from research experiment to clinical medicine.
Micromampulative skills opened the door to embryo biopsy for sexing by DNA amplification, detection of DNA fragmentation by TUNEL probe and embryo production by sperm injection, the first pregnancy after mtracyiopiasmic sperm injection (ICSI) was reported in 1992 (Palermo el al, 1992) and the birth of infant after transfer of anucleate donor oocyte cytoplasm was reported in 1997 (Cohen et al. 1997), The efficient freezing technique was developed and allowed to conserve oocyte, embryo and ovarian tissue for long time In 2004, the first woman in the world, who had parts of ovaries removed for freezing before beginning treafmenl for Hodgkm's lymphoma and reimplanted six years later, h;i-, given birth to a
Bui Xuan Nguyen el ai healthy baby girl (Donnez et ai. 2004).
After the birth of Louise Brown on 25 July 1978, Edwards and Steptoe established the Bourn Hall Clinic in Cambridge, the world's first centre for IVF therapy where the gynecologists and cell biologists around the worid were trained. Today, IVF technique spreads around the world as a refined, safe and effective therapy with 20-30% of fertilized eggs leading to the birth of child The number of babies bom using assisted reproductive technologies (ART) has reached an estimated total of 5 million; around 1,5 million of ART cycles are now performed globally each year, producing around 350,000
The application of embryo technology to animal breeding was one of the most far-reaching advances occurred early since the 80's decade. Efficient techniques of in vitro maturation and IVF, sperm and embryo sexing were established for all domestic species The preservation and transport of embryo were developed from an primitive experiment realized by Marden and Chang in 1952 when rabbit embryos were kept in the oviduct for a short air travel to successful cryopreservation and transfer in liquid nitrogen for unlimited time and distance (Whittingham et ai. 1972; Renard et ai, 1984) Embryo technology has developed at industrial level covered the genetic selection for almost all animal breeding station and farm in the developed countries.
Number of embryo transfer has estimated more than half of millions per year.
The SCNT has been used as a new promising tool for biodiversity conservation. Wells and co- workers reported in 1998 the first successful use of SCNT for preserving the last surviving cow of the Enderby Island cattle breed. The successful cloning of an adult from animal keeping in refrigerator for 16 years reported by Wakayama and colleagues (2008) proving the usefiilness of SCNT for the purpose of clonal expansion to restore endangered, or even extinct species such as Siberia Mammoth.
The use of SCNT as a new approach for gene transfer has a great impact on the production of transgenic animals. In comparison with transgenic animals from microinjection of DNA into pronuclei, the use of SCNT increases efficiency of production of transgenic farm animals to 100%. SCNT have applied successfiilly for Gene 'Pharming' entails the production of recombinant pharmaceutically active human proteins in the mammary gland or blood of
transgenic animals. SuccessfijI cloning pigs with the b-l,3-galactosyltransferase gene knocked out for xeno-transplantation was reported by different laboratories (Lai et ai, 2002).
Stem cell studies are currently under intense investigation in many laboratories to develop regenerative medicine (Yang el ai, 2007). The positive results of testing retinal cells derived from human embryonic stem cells for blind patients recently reported showing that the cells appear to be safe after being injected into the eyes and resulted in patient's visual improvements (Steven et a/., 2012).
Recently, Qian and co-workers (2012) used in vivo reprogramming as the new approach, which allowed the differentiation o f t h e cells in their native environment. Their results show that cardiac fibroblasts can be reprogrammed into cardiomyocyte-like cells and resulted in fiirther improvements in scar area and cardiac function in mice receiving local delivery of Gata4, Mef2c and Tbx5 (GMT) after coronary ligation.
Remarkable results were obtamed in research of using stem cells for reproductive restoration purpose.
In 2011 and 2012, a Kyoto University team led by stem cell biologist Mitinori Saitou reported successful production p f mouse sperm and oocytes from both embryonic stem cells and induced plunpotent stem cells; these sperm and oocytes were then used for IVF, leading to normal birth of mouse pups after transplantation into surrogate mothers (Hayashi era/., 2012).
The use of genetically engineering T-cells differentiated from stem cells for HIV and cancer treatment is a recent trend of research with the great potential and hope. The miracle story of Patients HIV-Free after Bone Marrow Transplants recently reported at XIX Intemational AIDS Conference (AIDS 2012) inspired scientists to look for new ways to vanquish HIV disease. In recent experimental work in animal model, Scott G. Kitchen group demonstrated for the first time that engineering stem cells are capable to form immune cells that target HIV for effecfive suppressing the virus in living tissues (Scott et ai, 2012).
The successful establishment of human embryonic stem cell (hESC) lines has raised high expectation for their future applications on safety and toxicological assessments in 628
Tgp chi Cong nghe Sinh hgc 10(4): 625-632, 2012 dmg discovery and drug development. Using stem cells to weed out drugs with dangerous side effecls before they reach the market could save the industry millions of dollars in wasted development costs.
Research and Development of embryo and stem cells at VAST
The basic technologies for embryo and stem cells research and development have been established at the Academy of Science and Technology of Vietnam (Nguyen, 2006), During the period 1978-2005, these studies had been carried out to develop embryo technologies, which were adapted to animals under tropical and non- standardized breeding conditions in Vietnam, The first successful embryo transfer was reported for rabbit in 1978, for cattle in 1986 and for goat in 1996 (Fig. IA; B; C). The development of original principle for embryo preservation by rapid freezing after partial dehydration at room temperature was successfully developed in both in VIVO as well as SCNT embryos (Nguyen el ai.
1984, 2000) and have been applying now for cryobanking of different genetic resources (Uoc et al., 2002; Hanh ei ai, 2007). In the collaboration
with BOMED-USA, INRA France, the first calves from IVF embryos and from IVF-sexing embryos were bom in 2002. The intercontinental IVF shipped embryos transfer was established for transfer of IVF-sexing embryo into local Zebu- yellow recipients resulted in the average calving rate of more than 50%. This technology was applied for production of cattle reproductors of selected quality (Nguyen et ai. 2004)
The study of somatic cell nuclear transfer has been carried out since 1998 as an approach for conservation of animal biodiversity. A model of interspecies nuclear transfer using bovine enucleated oocytes as universal host ooplasts for reprogramming of somatic cell nuclear was successfully developed for embryo production in Saola {Pseudoryx ngheiinhensis)-a new mammalian species discovered in Vietnam (Fig, ID, E) (Bui el ai. 2002, Trung et ai. 2008). In 2007, under support of BIODIVA project, an national and regional ex situ-in situ integration basing on ART technology as a new approach for active conservation of animal biodiversity was proposed for South East Asian countries (Nguyen et ai. 2007),
Figure 1. A First result of embryo transfer. 1978; 8. Goat frozen embryo transfer, 1996, C. First IVF-Sexing Twine calves, D S a o l a , 1 9 9 4 . E iSCNT Saola blastocyst: F Neuronal like Stc from HUMC
Bui Xuan Nguyen et ai Since 2005, studies on embryonic stem cell
technology were realized after receiving the formation at Stem Cells and Regeneration NIH Training Course, Pittsburgh, USA in 2004. During the years 2007-2011, under a national project supported by MOST, the production of neuronal and cardiac like stem cells (Fig. IF) by in vitro differentiation of human umbilical cord membrane had been carried out (Tran el ai, 20! I). The Japan- Vietnam project using stem cells and modem reproductive technique such as somatic nuclear transfer, embryo production by fiasion of cytoplasmic fragments for conservation of local porcine breeds with non-PERV and application for xeno- transplantation is underway (Uoc et ai. 2008; Q Thanh el ai. 2010; Linh cf A/., 2011).
CONCLUSION
In the 2 r ' ceniury, the development of new theories and instruments allows us to understand the cell activity at its smallest, most fundamental limits in length and in time such as nanoscale and femtosecond scale; ihe integration between informalic, biological and physique-chemical technologies will given the most importanl developmenls for biotechnology m general, embryo and stem cell technologies in particular.
Medicine and human health will enter a new age when the use of embryo and cell therapy has tremendous potential. The development of embryo and stem cell research and their applications is important for Vietnam, whose population would grow to more than 100 millions in the coming years and approximately 10% o f t h e population would be suffered from diabetic, 15% suffered from infertility The establishment of a research system as a platform for fundamental research, technological standardization and formation of experts-scientists in embryonic stem cells and stem cell research is the first need to face these requirements and challenge.
Acknowledgement: Wc thank VAST and Vietnam National Foundation for Science and Technology Development (NAFOSTED) for presiding financial help during paper preparation.
REFERENCES
Aldous Huxley Brave New World, First published by Chano & Windus, 1932
Briggs R, King TJ (1952) Transplantation of living nuclei from blastula cells into enucleated frogs eggs. Proc Natl AcadSci f;5A 38, 455-463.
Bui LC, X Vignon, E Campion, E Laloy, Y Lavengne, LV Ty, EX Nguyen, JP Renard (2002) Use of interspecific nuclear transfer to study the early embryonic development and nuclear activities ofthe endangered species Pseudoryx ngetmhensis (Saola). Thenogeiiology 57 427 ChangMC ('1959) FertihzaUon of rabbit ova in vitro.
Naturel84: 466-467,
Cohen R. Schimmel T, Levron J, Willadsen S (1997) Birth of infant after Iransfer of anucleate donor oocyte cytoplasm into recipient eggs Uncet, I9;350(9072),186-7.
Dang-Nguyen Quang Thanh, Nguyen Khac Tich, Bui Xuan Nguyen, Kazuhiro Kikuchi, Takashi Nagai (2010) Introduction of some Vietnamese indigenous pig breeds and their conservation hy using assisted reproductive techniques. J Rep Dev 56" 3 I -35
Donnez J, Dolmans MM, Demylle D, Jadoul P, Pirard C, Squifflet J, Martinez-Madrid B, van Langendonckt A (2004) ve birth after orthotopic transplantation of cryopreserved ovarian tissue, Li:(«cM364(9443), 1405-10.
Evans M, Kaufman M (1981) Establishment m culture of pluripotent cells from mouse embryos Nature 292 (5819)- 154-6
Gail R Martm (1981) Isolation of a plunpotent cell line from early mouse embryos cultured in medium conditioned by leralocarcinoma stem cells. Proc Nai Acad Sci USA 7&{\2y 7634-7638
Gurdon JB (1962) The developmental capacity of nuclei taken from intestinal epithelium cells of feeding tadpoles. J Embryo Exp Morphol 10: 622-40.
Jullien J, Astrand C, Szenker E, Garrett N, Almouzni G, Gurdon J (2012) HIRA dependent H3.3 deposition is required for transcriptional reprogramming following nuclear transfer to Xenopus oocytes. Epigenet Chromatin 5(1) 17
Hanh Nguyen Van, Qwan Xuan Huu, Nguyen Viet Linh, Nguyen Thi Men, Nguyen Thi Uoc, K Kukuchi, T Nagai, P Comizzoli, Bui Xuan Nguyen (2007) Conservation of endangered species semen. Genetics and Applications, special issue on Biodiva-VAST International Conference on «Reproductive biotechnologies for conservation of biodiversity and sustainable development », Hanoi, Dec, 17-18,2007,99-103.
Hayashi K. Ogushi S, Kunmolo K, Shimamolo S, Ohia H, Saitou M (2012) Offspring from oocytes derived from in vitro primordial germ cell-like cells in mice. Science 338(6109): 971-5
Heape W (1891) Preliminary note on the transplantation
Tap chi Cong nghe Sinh hoc 10(4): 625-632, 2012 and growth of mammalian ova within a uienn foster- mother. Proc Royal Soc B 48' 457-458,
Ul L, Kolber-Simonds D, Park KW, Cheong HT, Greenstem JL, Im GS, Samuel M, Bonk A, Rieke A, Day BN (2002a) Production of alpha- 1,3-gaIaclosyItransferase knockout pigs by nuclear transfer cloning. Science 295:
1089-92.
Linh NV, Kazuhiro Kikuchi, Michiko Nakai, Junko Noguchi, Hiroyuki Kaneko, Thanh Quang Dang-Nguyen, Naoki Maedomari, Bui Xuan Nguyen, Takashi Nagai, Nobom Manabe (2011) Improvement of porcine oocytes wilh low developmental ability after fusion of cytoplasmic fragments prepared by serial centrifugal ion, J Rep Dev 57{5):620-6.
Nguyen BX, Heyman Y, Renard JP (1984) Direct freezing of cattle embryos afler partial dehydration at room temperamre. Theriogenology 22,4,389-399
Nguyen BX, Sotomaru Y, Tani T, Kato Y, Tsunoda Y.
(2000). Efficient cryopreservation of bovine blastocysts derived from nuclear transfer with somatic cells using partial dehydration and vitrification. Theriogenology, 53:1439-1448
Nguyen BX (2004) The development of embryonic and cloning technology at Viemam Academy of Science and Technology IrsiAsian Biotechnology Conference. Ho Chi MinhAprii 13-17.
Nguyen Bui Xuan (2006) Current status and trends of animal reproductive biotechnology in Vietnam Embryo 'fTa.nsfe,'c'Htvi&\e,\.,Reprod Fen Developmeni 24(2) 5-10 Nguyen Bui Xuan (2007) Biotechnology aspect of active conservation and contribution lo sustainable development, VAST International Conference on "Reproductive biotechnologies for conservation of biodiversity and sustainable development": 17-18,.
Palermo G, Joris H, Devroey P, Van Sleirteghem AC (1992) Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte Lancet 340(8810)* 17-8 Qian L, Huang Y, Spencer CI, Foley A, Vedantham V, Liu L. Conway SJ, Fu JD, Srivaslava D (2012) In vivo reprogramming of murine cardiac fibroblasts into induced cardiomyocytes Nature 485'593-598
Renard JP, Bui-Xuan-Nguyen, Gamier V (1984) Two -step freezing of two -cell rabbit embryos after partial dehydration at room temperature. J Reprod Pert 71 573-580.
Ring KL, Tong LM, Balestra ME, Javier R. Andrews- Zwillmg Y, Li G. Walker D, Zhang WR, Kreitzer AC, Huang Y (2012) Direct reprogramming of mouse and human llbroblasis into multipotent neural stem cells with a single factor. Cell Slem Cell 11 (1): 100-9.
Schwartz SD, Hubschman JP, Hcilwell G, Franco- Cardenas V, Pan CK. Ostrick RM. Mickunas E. Gay R,
Klimanskaya I, Lanza R (2012) Embryonic stem cell tnals for macular degeneration, a preliminary report Lancet 379(9817): 713-20.
Scott G. Kitchen, Bemard R. Levin, Gregory Bnslo, Valerie Rezek, Sohn Kim Christian A gui I era-San do vai, Animugam Balamurugan, Otto O Yang, Jerome A Zack (2012) In Vivo Suppression of HIV by Antigen Specific T Cells Denved from Engineered Hematopoietic Stem Cells PLoS Pathogens 8(4)
Steptoe PC, Edwards RG (1978) Birth after the reimplantation of a human embryo Lancet 2(8085): 366, Stummann T C, Bremer S (2008) The possible impact of human embryonic stem cells on safety pharmacological and toxicological assessments in drug discovery and drug developmeni Curr Stem Cell Res Ther 3{2). 118-31, Takahashi K, Yamanaka S (2006) InducUon of plunpotent stem cells from mouse embryonic and adult fibroblast culmres by defined factors Cell \26 663-676, Thibault C, Dauzier L, Wintenberger S (1954) Emde cylologique de La fecondaiion m vitro dePoeuf de la lapme. C R Soc Biol (Pans) 148: 789-790.
Thomson James, Joseph Itskovitz-Eldor, Sander S.
Shapiro, Michelle A. Waknitz, Jennifer J Swiergiel, Vivienne S, Marshall, Jeflrey M Jones Embryonic Stem Cell Lines Derived from Human Blastocyst Science 282, 1145(1998)
Till JE, McCulloch EA (1961) A direct measurement of the radiation sensitivity of normal mouse bone marrow ceW%". Rad Res 14(2) 213-22.
Tran Thi Thom, Nguyen Thi Men, Nguyen Thi Uoc, Bui Linh Chl, Bui Xuan Nguyen (2010) Effect of difTerenlialion factors on the induction of human umbilical cord matnx slem cell into neural progenitor cells.
Procceedings of the international conference of ARB, 2010. Malaysia: 1-5.
Tmng Thanh Nguyen, Aniskin VM, Gerbault-Seureau M, Planlon H, Bui Xuan Nguyen, JP Renard, Alexandre Hassanin, Vitaly Volobouev (2008) Phylogenetic position of the saola (Pseudoryx nghetinhensis) inferred from cytogenetic analysis of eleven species of Bovidae Cylogenel Genome Res 122. (DOI: 10.1159/000151315) Uoc NT, Chl BL, Duc NH, Ty LV, Keo N, Tuoc D, Thanh NT, Hanh NV, E, Laloy, Renard JP, Nguyen BX (2002) Effect of tissue sampling conditions on the In Vitro mulliplication and reprogramming potentail uf somatic cells obtained from different specimens of the Saola (Speudoryx Nghetinhensis) species Tliern/genulogv 57:437
Uoc Nguyen Thi. Le Van Ty, Nguyen Huu Duc, Bui Linh Chi, Nguyen Trung Thanh, Nguyen Viet Linh. Nguyen Van Hanh, Quan Xuan Huu. Nguyen Thuy Anh, Hoang Nghia Son, Duong Dmh t>ong, Bui Xuan Nguyen , (2003).
631
Bui Xuan Nguyen et ai Production of dairy tropical calves by trasfer of IVF and
sexed embryos. Proceedings of Vietnam National Conference of Biotechnology, 717-719
Uoc Nguyen Thi, Nguyen Viet Linh, Nguyen Van Hanh, Quan Xuan Huu, Dang Nguyen Quang Thanh, Nguyen Thi Men, Tran Thi Thom, Bui Linh Chi, Nguyen Trang Thanh, Duong Dinh Long, Nguyen Khac Tich, Phan Ngoc Minh and Bui Xuan Nguyen (2008) Production of ative mimpig embryo by complex of in vitro techniques and somafic cloning. J Biotechnol 6(4A). 625-635, Vatakis DN, RC Koya, CC Nixon, L Wei, SG Kim. P Avancena, G Bristol. D Baltimore, DB Kohn, A Ribas, CG Radu, 2 Galic, JA Zack (2011) PNAS Plus Antitumor activity from anUgen-specific CDS T cells generated in VIVO from genetically engineered human hematopoietic stem cells. Proceedings of the National Academy of Sciences 108(51).
Wakayama S, Ohta H, Hikichi T, Mizutani E, Iwaki T,
Kanagawa O, Wakayama T (2008) Production of healthy cloned mice from bodies frozen at -20 degrees C for 16 years Proc Natl Acad Sci USA 105(45): 17318-22.
Wells RJ, Misica PM, Tervit HR (1999) Production of cloned calves following nuclear transfer with cultured adult mural granulosa cells Biol Reprod 60- 996-1005.
Wilmut I. Schmeke AE, McWhir J, Kind AJ, Campbell BCHS (1997) Viable offspnng derived from fetal and adult mammalian cells Naiure 3%5:810-S\3.
Whittingham DG, Leibo SP, Mazur P (1972) Survival of mouse embryos frozen to-196 and-269<'C. Science 178:
411-414.
Yang Xiangzhong, Sadie L Smith, X Cindy Tian, Hams A Lewin, Jean-Paul Renard, Teruhiko Wakayama (2007) Nuclear reprogramming of cloned embryos and its implications for therapeutic cloning Nai Genet 39,295-302
CONG NGHE PHOI VA TE BAO GOC THE KY XXI
Bui Xuan Nguyen', Nguyen Thi U"orc, Biii Linh Chi, Nguyin Trung Thanh, Nguyen Viet Linh, Dang Nguyin Quang Thanh, Nguyin Van Hanh, Nguyen Thi Men, Tran Thi Thom, Nguyen Thi Hiep, N g u y i n Thi Nhung, Nguyen Thi Hong
Vien Cong ngh^ sinh hgc. Vien Khoa hgc vd Cong nghe Viet Nam
TOM TAT
The ky 21 se chimg kien nhuTig tac dong lo Ion ciia cong nghe sinh hoc Ien hoal dong song ciia loai nguoi, Viec trao giai Nobel smh ly - y hoc nam 2012 cho cac nha khoa hoc nghien ciru ve nhan ban vo tinh va te hao goc danh dau mpl thai ky hung no cac y tuong nghien cim va iing dung moi trong ITnh virc cong nghe ph6i va te bao goc, Trong bai long hop nay chung loi se trinh bay lugtc sir cac tu tucmg khoa hoc va tien bo ky thuat lien quan cong nghe phoi va te bao goc nhu ly thuyet ve kha nang ma hoa lai cua Spemann (1932), su ra doi ciia Ihu tinh ong nghiem tren nguoi vao nSm 1978, viec sir dung thanh cong ky ihuat cay nhan de t^o dong vat nhan ban v6 tinh vao nam 1197, sir phat hien le bao goc tiiy song vao nam 1963 va va te bao da nang nhan t^o vao nam 2006; dong thai de cap hien tr^ng nghien ciiu, anh huong va trien vpng ciia cac phat minh nay Ien hoat dpng chan nuoi, bao ton da dang sinh hpc, y hpc va sire khoe con nguoi Bai bao ciing de cgp van tat cac kel qua nghien ciju ve cay chuyen phoi, san xuat phoi bJng cac cong nghe thu Unh ong nghiem va nhan ban vo tinh, dong lanh va chon gioi tinh phoi cac loai dong vat nuoi va dong vat hoang da, nghien ciru bao quan va biet hoa te bao goc nguoi da duoc lien hanh tai Vien Han lam khoa hoc va cong nghe Viet Nam, nhu chx vii thach thuc doi voi viec phal trien cac cong nghe nay a Viet Nam trong boi canh hien tai.
Tir khoa: Cong nghe. phoi. te bdo goc. The ky XXI. Viin Cong nghe sinh hoc
^Authorfor correspondence: Tel. +84-4-7>15h29G2; Fax: +84-4-l>19\-263l\ E-rnaiL buixn5@smaiirom 632