• Journal of Embryo Transfer
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• v.20 no.3
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• pp.239-253
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• 2005

Serum concentrations of Hanwoo steers and bulls as possible indicators of beef quality were analyzed to estimate their correlations with carcass traits. Blood samples were taken 2 months and right before shipping to abattoir and at the time of slaughter. And phenotypic correlation coefficients between serum concentrations and carcass traits were estimated. Beef yield index of steers was positively correlated with serum concentrations of total Protein (0.23), albumin (0.26), and calcium (0.31). But it was negatively co..elated with BUN (-0.30). Loin eye area was positively correlated with BUN (0.17) or with globulin (0.16). Back fat thickness was positively correlated with BUN (0.42) and inorganic phosphorus (0.20) being negatively correlated with total protein (-0.23), albumin (-0.33) and calcium (-0.33). Marbling score in the scale of 1 (scarcely marbled) through 9 (extremely marbled) was positively correlated with BUN (0.28) and negatively with IGF-I and calcium concentrations. Phenotype correlation coefficient of loin eye area with total protein concentration in the serum taken from steers right before shipment was estimated to be -0.16 and that with BUN was estimated to be -0.15. Serum concentrations of IGF, glucose, creatinine and on organic phosphorus from steers measured right before shipment were negatively correlated with respective correlation coefficient estimates as -0.21, -0.21, -0.19 and -0.18. Marbling score was negatively co..elated with serum creatinine (-0.16) measured at that time. Beef yield index of steers was positively correlated (0.31) with age adjusted calcium concentration in the serum taken at the time of slaughter. Correlation between body weight and BUN at slaughter was 0.17 At slaughter, loin eye area was negatively correlated with albumin (-0.19) and back fat thickness was also negatively correlated with age adjusted calcium concentration (-0.38). Marbling was negatively correlated with age adjusted calcium concentration(-0.17). Serum concentrations of testosterone, calcium and inorganic phosphorus taken in 2 months before slaughter were negatively but highly correlated with yield index(0.71, 0.67 and -0.71), respectively. Body weight at slaughter was positively was negatively correlated (0.67) with calcium level while dressing percentage was negatively (-0.69) correlated with serum glucose concentration, 2 months prior to slaughter. Correlation coefficients between back fat thickness and cortisol, between back fat thickness and inorganic phosphate were both positive (0.29 and 0.69). Marbling score was negatively correlated with creatinine (-0.81) and positively with BUN (0.87). Body weight loss during shipping was positively correlated with albumin and inorganic phosphate (0.77, 0.83). Yield index of bulls was positively correlated with serum testosterone concentration (0.66). Dressing percentage was positively and highly correlated with globulin (0.73). Back fat thickness of bulls, however, was negatively correlated with testosterone (-0.60). Loin eye area of bull carcasses was positively correlated with testosterone (0.40). Mar-blaine was negatively co..elated with creatinine (-0.55). Yield index of bulls and age adjusted HDLC concentration at slaughter was negatively correlated (-0.71). Dressing percentage of bulls was positively and highly correlated with globulin concentration (0.70). Back fat thickness was also positively correlated with HDLC (0.69) in the serum taken at slaughter. Correlation coefficients between carcass weight and triglyceride, between loin eye are and testosterone and between marbling score and creatinine or glucose were 0.51, -0.91 and -0.58, respectively.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.279-287
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• 2005

This study was carried out to investigate resumption of ovarian cyclicity and effect of BCS on estrous response by treatment of $PGF_2a+PGF_2a+CIDR$ program on day 40 postpartum in lactating dairy cow. First $PGF_2a$ was given on day 40 postpartum, second $PGF_2a$ was given 14 days apart to cows not-responded to 1st $PGF_2a$ and then CIDR was inserted for 7 days after 5 days in cows not-responded to 2nd $PGF_2a$. The $42.9\%$ of the cows showed more than 1 ng/mL milk progesterone concentration within 10 to 30 days postpartum. About $19\%$ of the cows exhibited more than 1 ng/mL milk pro-gesterone concentration between 31 to 50 days postpartum. However $38.1\%$ of the cows have not shown more than 1 ng/mL milk progesterone up to 50 days postpartum. Estrous response to the treatment of 1 st $PGF_2a$ and 2nd $PGF_2a$ was $47.5\%$ and $52.4\%$, respectively. Combination of 1 st $PGF_2a$ and 2nd $PGF_2a$ was $75\%$ and combination of 1st $PGF_2a$+2nd $PGF_2a$+CIDR was $87.5\%$. Estrous response to the treatment of $PGF_2a+PGF_2a$ program was $61.5\%$ in cows with less than 2.50 BCS and $81.5\%$ in cows with 2.75${\~}$3.50 BCS. Estrous response to the treatment of CIDR was $40\%$ in cows with less than 2.50 BCS and $80\%$ in cows with 2.75${\~}$3.50 BCS. Estrous response to the treatment of PPC on day 40 postpartum was $76.9\%$ in cows with less than 2.50 BCS and $96.3\%$ in cows with 2.75${\~}$3.50 BCS.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.109-119
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• 2006

This study was conducted to examine relationship between vaginal cytology and reproductive hormones during the estrous cycle and to provide basic data to estimate for ovulation time and optimal mating time in 6 beagle dogs The duration of proestrus, estrus and diestrus were $8.5{\pm}1.4,\;10.0{\pm}1.4\;and\;54.0{\pm}2.8$ days at pregnant respectively, and $7.9{\pm}2.1,\;9.5{\pm}0.7\;and\;62.0{\pm}11.3$ days at non-pregnant respectively. The duration of interestrous intervals were $246.2{\pm}24.5$ days at pregnancy, and $175.3{\pm}34.5$ days at non-pregnancy. The duration of interestrous intervals at pregnancy was longer than that of non-pregnancy. A characteristic features of vaginal cytology during the estrous cycle were the high proportion of superficial cell, anuclear cell and erythrocyte in proestrus and estrus, parabasal cell, small intermediate cell and leukocyte in diestrus, and parabasal cell and small intermediate cell in anestrus, respectively. Cornification index (CI) in proestrus and estrus were significantly higher than that of CI in diestrus and anestrus. Plasma progesterone concentration was below 1.0 ng/ml at the first day of vulval bleeding at pregnancy and non-pregnancy, and then it was above 2.0 ng/ml at Day -2 in all bitches. When plasma progesterone concentration was first increased above 4.0 ng/ml, it was the second day after the first day of male acceptance. Plasma progesterone concentration showed above 40 ng/ml on Day $20{\sim}22$ in all bitches, and then it was gradually decreased until Day 35. Plasma progesterone concentration at pregnancy was higher than that of non-pregnancy from Day 35 to Day 63. Plasma estradiol-$17\;{\beta}$ concentration was above 9.0 pg/ml at the first day of vulval bleeding, and it showed 26.4 pg/ml on Day -2. When it was timed from the first day of male acceptance (Day 0), plasma estradiol-$17{\beta}$ concentration showed a peak on Day 0 and plasma progesterone concentration was first increased above 4.0 ng/ml on Day 2 which was the third day after plasma estradiol-$17{\beta}$ peak. CI was first increased above 80 and 90% on Day -1 and Day 1, respectively. CI was maintained above 80% from Day -1 to Day 8 (10 days) and above 90% from Day 1 to Day 6 (6 days), respectively. CI was maintained above 80% from Day 0 to Day 8 (9 days) and above 90% from Day 1 to Day 6 (6 days), respectively. Plasma progesterone concentration was first increased above 4.0 ng/ml on the second day after the day which CI was first increased above 90%. In conclusion, beagle bitches ovulated on the second day after the day which CI was first increased above 90% and on the day which plasma progesterone concentration was first increased 4.0 ng/ml, and it was estimated that the optimal mating time was the day which the second day after CI was first increased above 90% and plasma concentration was between $2{\sim}25ng/ml$. The measurement of plasma progesterone was used to determine of and accurate ovulation time and the optimal mating time, but vaginal cytology is low-priced and simple method to estimate estrous cycle, optimal mating time and ovulation time.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.177-184
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• 2005

This study was conducted to investigate the effects of recombinant bovine somatotropin (rbST) injection on conception and parturition rates in normal or repeat breeding Hanwoo. We treated 462 cows containing 79 repeat-breeding cows of multiparous and allocating 5 treatment groups. Treatment 1 (T1) was injection of 2ml saline (for pseudo treatment), T2 was one injection of rbST 250mg into the tailhead region at the estrus, T3 was twice injection of rbST 250mg both at the time of insemination and again 10 to 14 day later, T4 was once injection of rbST 500mg at insemination and T5 was twice injection of 500mg rbST both at the time of insemination and again 10 to 14day later respectively. In rbST treated groups, timed artificial inseminations (TAI) were performed fellowing estrus synchronization. 100 us GnRH was injected into the scapula region on Day 0, 25mg $PGF_2{\alpha}$ was injected on Day 7 for degeneration of corpus luteum (CL) and 100ug GnRH was injected for inducing the synchronization. The results are as fellows; When normal Hanwoo were inseminated once with rbST administration, the pregnancy rate of T2 $(67.5\pm18.48\%)$ were higher than control $(52.4\pm9.72\%)$, while the pregnancy rate of T4 $(63.3\pm5.77\%)$ were significantly higher (p.<0.05) than control $(39.3\pm12.89\%)$ in repeat breeder Hanwoo. The parturition rates of normal Hanwoo were no differences among the treatments but were significant different in repeat breeder Hanwoo (p<0.05). When the estrous was induced by Ovsynch and inseminated once with rbST administration, the pregnancy rates of T2 was $12.5\%$ higher than control in normal Hanwoo, T4 $(80.0\%)$ was highest among the treatments (p<0.05) in repeat breeder Hanwoo. When normal Hanwoo were inseminated once with rbST administration, the pregnant period was $282.7\~284.8$ days and the body weight was $25.1\~25.9kg$, there were no difference among the treatments. The ratio of sex was almost same without T4 (male vs. female=18:9). In repeat breeder Hanwoo, pregnant period was 280.4~289.3 day and body weight was $23.0\~26.6kg$, it had no difference among the treatments. The sex ratio were similar to normal Hanwoo except T4 (M : F=2 : 8). In conclusion, the pregnancy and parturition rate by once insemination could be improved by the administration of rbST 250mg in normal Hanwoo or 500mg in repeat breeder Hawoo.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.1
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• pp.49-56
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• 2010

Objective: To investigate assisted reproductive technology (ART) outcomes in women with WHO class I anovulation compared with control group. Design: Retrospective case-control study. Methods: Twenty-three infertile women with hypogonadotropic hypogonadism (H-H) who undertook ART procedure from August 2003 to January 2009 were enrolled in this study. A total of 59 cycles (H-H group) were included; Intra-uterine insemination with super-ovulation (SO-IUI, 32 cycles), in vitro fertilization with fresh embryo transfer (IVF-ET, 18 cycles) and subsequent frozenthawed embryo transfer (FET, 9 cycles). Age and BMI matched 146 cycles of infertile women were collected as control group; 64 cycles of unexplained infertile women for SO-IUI and 54 cycles of IVF-ET and 28 cycles of FET with tubal factor. We compared ART and pregnancy outcomes such as clinical pregnancy rate (CPR), clinical abortion rate (CAR), and live birth rate (LBR) between the two groups. Results: There was no difference in the mean age ($32.7{\pm}3.3$ vs. $32.6{\pm}2.7$ yrs) and BMI ($21.0{\pm}3.1$ vs. $20.8{\pm}3.1kg/m^2$) between two groups. Mean levels of basal LH, FSH, and $E_2$ in H-H group were $0.62{\pm}0.35$ mIU/ml, $2.60{\pm}2.30$ mIU/ml and $10.1{\pm}8.2$ pg/ml, respectively. For ovarian stimulation, H-H group needed higher total amount of gonadotropin injected and longer duration for ovarian stimulation (p<0.001). In SO-IUI cycles, there was no significant difference of CPR, CAR, and LBR between the two groups. In IVF-ET treatment, H-H group presented higher mean $E_2$ level on hCG day ($3104.8{\pm}1020.2$ pg/ml vs. $1878.3{\pm}1197.7$ pg/ml, p<0.001) with lower CPR (16.7 vs. 37.0%, p=0.11) and LBR (5.6 vs. 33.3%, p=0.02) and higher CAR (66.7 vs. 10.0%, p=0.02) compared with the control group. However, subsequent FET cycles showed no significant difference of CPR, CAR, and LBR between the two groups. Conclusion: H-H patients need higher dosage of gonadotropin and longer duration for ovarian stimulation compared with the control groups. Significantly poor pregnancy outcomes in IVF-ET cycles of H-H group may be due to detrimental endometrial factors caused by higher $E_2$ level and the absence of previous hormonal exposure on endometrium.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.1
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• pp.29-39
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• 2004

Objectives: This study was performed to evaluate the laboratory system for successful PGD using fluorescence in situ hybridization (FISH) and the clinical outcome of PGD cycles in five years experiences. Methods: A total of 181 PGD-FISH cycles of 106 couples were performed, and diagnosed chromosome normality in the preimplantation embryos. The laboratory and clinical data were classified by the following optimization steps, and statistically analyzed. Phase I: Blastomere biopsy with two kinds of pipettes, removal of cytoplasmic proteins without treatment of pepsin and culture of biopsied embryos with single medium; Phase II: Blatomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with single medium; Phase III: Blastomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with sequential media. Results: A total of 3, 209 oocytes were collected, and 83.8% (2, 212/2, 640) of fertilization rate was obtained by ICSI procedure. The successful blastomere biopsies were accomplished in 98.6% (2, 043/2, 071) of embryos, and the successful diagnosis rate of FISH was 94.7% (1, 935/ 2, 043) of blastomeres from overall data. Embryo transfers with normal embryos were conducted in 93.9% (170/181) of started cycles. There was no difference in the successful rate of biopsy and diagnosis among Phase I, II and III. However, the pregnancy rate per embryo transfer of Phase III (38.8%, 26/67) was significantly (p<0.05) higher than those of Phase I (13.9%, 5/36) and Phase II (14.9%, 10/67). Conclusions: The laboratory optimization and experience for the PGD with FISH procedure can increase the pregnancy rate to 38.8% in the human IVF-ET program. Our facility of PGD with FISH provides the great possibility to get a normal pregnancy for the concerned couples by chromosomal aberrations.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.4
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• pp.253-260
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• 2004

Objectives: Klinefelter syndrome is the most common genetic cause of male infertility and presents with 47, XXY mainly or 46, XX/47, XXY mosaicism. It is characterized by hypogonadism and azoospermia due to testicular failure, however, sporadic cases of natural pregnancies have been reported. With the development of testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI), sperm can be retrieved successfully and ART is applied in these patients for pregnancy. It has been suggested that the risk of chromosome aneuploidy for both sex chromosome and autosome is increased in the sperms from 47, XXY germ cells. Considering the risk for chromosomal aneuploidy in the offspring, preimplantation genetic diagnosis (PGD) could be applied as a safe and more effective treatment option in Klinefelter syndrome. The aim of this study is to assess the outcome of PGD cycles by using FISH for sex chromosome and autosome in patients with Klinefelter syndrome. Materials and Methods: From Jan. 2001 to Dec. 2003, PGD was attempted in 8 cases of Klinefelter syndrome but TESE was failed to retrieve sperm in the 3 cases, therefore PGD was performed in 8 cycles of 5 cases (four 47, XXY and one 46, XY/47, XXY mosaicism). In one case, ejaculated sperm was used and in 4 cases, TESE sperm was used for ICSI. After fertilization, blastomere biopsy was performed in $6{\sim}7$ cell stage embryo and the chromosome aneuploidy was diagnosed by using FISH with CEP probes for chromosome X, Y and 17 or 18. Results: A total of 127 oocytes were retrieved and ICSI was performed in 113 mature oocytes. The fertilization rate was $65.3{\pm}6.0%$ (mean$\pm$SEM) and 76 embryos were obtained. Blastomere biopsy was performed in 61 developing embryos and FISH analysis was successful in 95.1% of the biopsied blastomeres (58/61). The rate of balanced embryos for chromosome X, Y and 17 or 18 was $39.7{\pm}6.9%$. The rate of aneuploidy for sex chromosome (X and Y) was $45.9{\pm}5.3%$ and $43.2{\pm}5.8%$ for chromosome 17 or 18, respectively. Embryo transfer was performed in all 8 cycles and mean number of transferred embryos was $2.5{\pm}0.5$. In 2 cases, clinical pregnancies were obtained and normal 46, XX and 46, XY karyotypes were confirmed by amniocentesis, respectively. Healthy male and female babies were delivered uneventfully at term. Conclusion: The patients with Klinefelter syndrome can benefit from ART with TESE and ICSI. Considering the risk of aneuploidy for both sex chromosome and autosome in the sperms and embryos of Klinefelter syndrome, PGD could be offered as safe and more effective treatment option.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.3
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• pp.207-215
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• 2003

Objective: To compare the pregnancy outcomes after in vitro fertilization with intracytoplasmic sperm injection (IVF-ICSI) between obstrucvtive and non-obstrucvtive azoospermia. Methods: From January 1994 to December 2002, 524 patients with obstructive azoospermia (886 cycles) and 163 patients with non-obstructive azoospermia (277 cycles) were included in this study. Microsurgical epididymal sperm aspiration (MESA) or testicular sperm extraction (TESE) in obstructive azoospermia and TESE in non-obstructive azoospermia were perfomed to retrieve sperm, which was used for ICSI and then fertilized embryos were transferred. The results of ICSI - fertlization rate (FR), clinical pregnancy rate (CPR), clinical abortion rate (CAR) and delivery rate (DR) - were statistically analysed in obstructive versus non-obstructive azoospermia. Results: There were no differences in the number of retrieved oocytes, injected oocytes for ICSI and oocyte maturation rate. FR was significantly higher in obstructive than non-obstructive azoospermia (71.7% vs. 61.1%, p<0.001). There was no difference in CPR per embryo transfer cycle. After pregnancy was established, however, CAR was significantly higher in non-obstructive than obstructive azoospermia (25.6% vs. 12.5%, p=0.004). DR per clinical pregnancy cycle was significantly higher in obstructive than non-obstructive azoospermia (78.0% vs. 64.4%, p=0.012). In the karyotype ananlysis of abortus, abnormal karyotypes were found in 75.0% (6/8) of obstructive and 55.6% (5/9) of non-obstructive azoospermia. Conclusion: Our data show significantly higher FR in obstructive than non-obstructive azoospermia. Though there was no differrence in CPR, CAR was significantly higher in non-obstructive than obstructive azoospermia. The abortion may be related to the abnormal karyotype of embryo, but further investigations are necessary to elucidate the cause of clinical abortion in azoospermia.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.83-93
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• 1997

Although IVF-ET is widely applied in the treatment of couples with male factor infertility, it may fail in many infertile couples with normal semen parameters, and certain couples cannot be accepted for standard IVF-ET due to unfertilization or extremely low fertilization rate of oocytes. Recently, several procedures of microassisted fertilization (MAF) using micromanipulation have been introduced, and pregnancies and births have been obtained after partial zona dissection (PZD), subzonal insertion (SUZI), and intracytoplasmic sperm injection (ICSI). This clinical study was performed to develop and establish ICSI as an effective procedure of MAF in infertile couples who could not undergo standard IVF-ET repetitively because of failure in fertilization or extremely low fertilization rate of oocytes with the conventional fertilization technique in the previous IVF-ET cycles. From March, 1995 to May, 1996, 27 cycles of IVF-ET with ICSI in 19 infertile patients were included in study group, and the outcomes of ICSI were analyzed according to fertilization rate, cumulative embryo score (CES), and pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation (COH) was $10.50{\pm}6.13$ in 30 previous cycles, and $10.57{\pm}5.53$ in 27 ICSI cycles. In ICSI cycles, the number of oocytes optimal for ICSI procedure was $7.89{\pm}4.30$, and the fertilization rate of $67.9{\pm}20.2%$ could be obtained after ICSI. The number of embryos transferred was $1.43{\pm}2.40$ in previous cycles, and $4.36{\pm}1.77$ with the mean CES of $41.8{\pm}27.4$ in ICSI cycles. In ICSI cycles, the overall pregnancy rate was 29.6% (8/27) per cycle and 42.1% (8/19) per patient with the clinical pregnancy rate of 22.2% (6/27) per cycle and 31.6% (6/19) per patient. In conclusion, MAF of human oocytes with ICSI is a promising fertilization method for IVF-ET patients, especially with the past history of failure in fertilization or low fertilization rate of oocytes in the previous IVF-ET cycles, and ICSI using micromanipulation procedures applied to human oocytes will provide a range of novel techniques which may dramatically improve the pregnancy rate in IVF-ET program and contribute much to effective management of infertile couples.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.423-428
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• 2000

In 1997 when cloned sheep Dolly and soon after Polly were born, it had become head-line news because in the former the nucleus that gave rise to the lamb came from cells of six-year-old adult sheep and in the latter case a foreign gene was inserted into the donor nucleus to make the cloned sheep produce human protein, factor IX, in e milk. In the last few years, once the realm of science fiction, cloned mammals especially in livestock have become almost commonplace. What the press accounts often fail to convey, however, is that behind every success lie hundreds of failures. Many of the nuclear-transferred egg cells fail to undergo normal cell divisions. Even when an embryo does successfully implant in the womb, pregnancy often ends in miscarriage. A significant fraction of the animals that are born die shortly after birth and some of those that survived have serious developmental abnormalities. Efficiency remains at less than one % out of some hundred attempts to clone an animal. These facts show that something is fundamentally wrong and enormous hurdles must be overcome before cloning becomes practical. Cloning researchers now tent to put aside their effort to create live animals in order to probe the fundamental questions on cell biology including stem cells, the questions of whether the hereditary material in the nucleus of each cell remains intact throughout development, and how transferred nucleus is reprogrammed exactly like the zygotic nucleus. Stem cells are defined as those cells which can divide to produce a daughter cell like themselves (self-renewal) as well as a daughter cell that will give rise to specific differentiated cells (cell-differentiation). Multicellular organisms are formed from a single totipotent stem cell commonly called fertilized egg or zygote. As this cell and its progeny undergo cell divisions the potency of the stem cells in each tissue and organ become gradually restricted in the order of totipotent, pluripotent, and multipotent. The differentiation potential of multipotent stem cells in each tissue has been thought to be limited to cell lineages present in the organ from which they were derived. Recent studies, however, revealed that multipotent stem cells derived from adult tissues have much wider differentiation potential than was previously thought. These cells can differentiate into developmentally unrelated cell types, such as nerve stem cell into blood cells or muscle stem cell into brain cells. Neural stem cells isolated from the adult forebrain were recently shown to be capable of repopulating the hematopoietic system and produce blood cells in irradiated condition. In plants although the term$\boxDr$ stem cell$\boxUl$is not used, some cells in the second layer of tunica at the apical meristem of shoot, some nucellar cells surrounding the embryo sac, and initial cells of adventive buds are considered to be equivalent to the totipotent stem cells of mammals. The telomere ends of linear eukaryotic chromosomes cannot be replicated because the RNA primer at the end of a completed lagging strand cannot be replaced with DNA, causing 5' end gap. A chromosome would be shortened by the length of RNA primer with every cycle of DNA replication and cell division. Essential genes located near the ends of chromosomes would inevitably be deleted by end-shortening, thereby killing the descendants of the original cells. Telomeric DNA has an unusual sequence consisting of up to 1,000 or more tandem repeat of a simple sequence. For example, chromosome of mammal including human has the repeating telomeric sequence of TTAGGG and that of higher plant is TTTAGGG. This non-genic tandem repeat prevents the death of cell despite the continued shortening of chromosome length. In contrast with the somatic cells germ line cells have the mechanism to fill-up the 5' end gap of telomere, thus maintaining the original length of chromosome. Cem line cells exhibit active enzyme telomerase which functions to maintain the stable length of telomere. Some of the cloned animals are reported prematurely getting old. It has to be ascertained whether the multipotent stem cells in the tissues of adult mammals have the original telomeres or shortened telomeres.