Fertility preservation in women with cancer / 대한생식의학회지

Fertility preservation (FP) is an effort to retain the fertility of cancer patients, and as an emerging discipline, it plays a central role in cancer care. Because of improvement in diagnostic and therapeutic strategies, an increasingly large number of patients are surviving with cancer. FP specialists should make an effort to spread the significance of FP among reproductive women with cancer and provide appropriate education both for associated physicians and for cancer patients who wish to preserve their fertility. Physicians who take part in the initial diagnosis and management of cancer should consider the importance of early referral of young cancer patients to FP specialists and take care of those patients by providing timely information and appropriate counseling. Individualized treatment strategies should be delivered depending on the patient's situation with appropriate team approach.

Ovarian Hyper-Response to Administration of an GnRH-Agonist Without Gonadotropins

Several case reports have indicated that a small subgroup of patients may develop ovarian hyperstimulation following the administration of gonadotropin-releasing hormone agonists (GnRHa) without gonadotropins. However, since only few such cases have been published, it is unclear what course to follow in subsequent cycles after ovarian hyperstimulation in the first cycle using only GnRHa. A 33-yr-old woman was referred to in vitro fertilization for oocyte donation. A depot preparation (3.75 mg) of tryptorelin without gonadotropins induced ovarian multifollicular enlargement with high estradiol level, and was followed by human chorionic gonadotropin administration and oocyte retrieval. In a subsequent cycle of the same patient, a low dose of tryptorelin (0.05 mg) did not induce ovarian hyperstimulation, and resulted in clinical pregnancy. This report shows potential management of ovarian hyperstimulation following the administration of GnRHa without gonadotropins.

Barium Stimulates the Expression of Osteogenic Genes in Human Mesenchymal Stem Cells into Osteoblasts in vitro / 대한폐경학회지

OBJECTIVES: The purpose of this study was to determine the effect of barium on gene expression in differentiation of human mesenchymal stem cells into osteoblasts in vitro. METHODS: Human bone marrow stem cells were cultured for 0~14 days in osteogenic differentiation medium with strontium chloride (SrCl2) and barium chloride (BaCl2). Alkaline phosphatase (ALP) activity staining was the method selected for measuring osteoblast differentiation. Total ribonucleic acid (RNA) was extracted after 1, 3, 7, and 14 days, and analysis of runt-related transcription factor 2/core-binding factor alpha 1 (Runx2/Cbfa1), bone morphogenetic protein-2 (BMP-2), and bone sialoprotein (BSP) gene expression was performed by real-time reverse transcriptase (RT)-polymerase chain reaction (PCR). RESULTS: Barium and strontium had a superior enhancing effect on cell proliferation when compared to cells cultured in media without strontium or barium. BaCl2 produced a 2-fold increase in the expression of Runx2/Cbfa1 at 14 days. SrCl2 (0.1~0.3 mM) produced a 2-fold increase in the expression of Runx2/Cbfa1 at 14 days. Barium produced a 1.5-fold increase in the expression of BMP-2 on days 1 or 3. Expression of BSP was increased 1.5~1.7- and 2-fold on days 1 and 14 by barium and strontium, respectively. CONCLUSION: Barium-like strontium is considered one of the important factors in inducing mesenchymal stem cells to differentiate into osteoblasts with further enhancement on bone formation.

Microarray Analysis of Gene Expression During Differentiation of Human Mesenchymal Stem Cells Treated with Vitamin E in vitro into Osteoblasts

OBJECTIVE: Supplementation with vitamin E is able to protect bone against free radical-induced elevation of bone-resorbing cytokines. We examined gene expression by microarray analysis during the differentiation of human mesenchymal stem cells treated with vitamin E into osteoblasts in vitro. METHODS: Human bone marrow stem cells were cultured in osteogenic differentiation medium and vitamin E was added. A colorimetric immunoassay for the quantification of cell proliferation was used to measure osteoblast differentiation. Gene expression was analyzed using a microarray technique. We also used a real time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: It was found that vitamin E enhanced cell proliferation when compared to cells cultured in media without vitamin E. We focused on 68 genes which are related to osteogenesis and osteoclastogenesis. Alkaline phosphatase, transforming growth factor-beta 1, fibroblast growth factor receptor 1, matrix metalloproteinase 2, muscle segment homeobox 2, bone morphogenetic protein 1, biglycan, vascular endothelial growth factor B, dentin sialophosphoprotein, cartilage oligomeric matrix protein, runt-related transcription factor 2, fibroblast growth factor receptor 3, and SMAD2 were upregulated > 2-fold compared to the control. Conversely, osteopetrosis-associated transmembrane protein 1, microphthalmia-associated transcription factor, and epidermal growth factor receptor were downregulated > 2-fold compared to the control. Vitamin E produced a 1.5-fold increase in the expression of runt-related transcription factor 2 and transforming growth factor-beta 1 as determined by real time RT-PCR. CONCLUSION: Vitamin E had a positive effect on the gene expressions regarding osteogenic differentiation of mesenchymal stem cells.

The Association between Components of the Metabolic Syndrome and Bone Mineral Density

OBJECTIVE: The aim of this study was to evaluate the association between the components of the metabolic syndrome (MS) and bone mineral density (BMD). METHODS: We conducted a retrospective cross-sectional analysis for 128 men (mean age 50.4 +/- 8.3 years) and 391 women (mean age 46.9 +/- 11.6 years) in Korea University Guro Hospital, Korea. Height (cm), weight (kg), waist circumference (cm), and blood pressure (mmHg) were measured. Fasting plasma glucose (FPG), high-density lipoprotein cholesterol (HDL-C), and triglycerides were measured. BMD at the total hip and lumbar spine was measured by dual X-ray densitometry. All participants completed a standardized questionnaire including medical history, smoking history, alcohol consumption, physical activity and menopausal status. RESULTS: Women with MS had lower BMD at lumbar and total hip sites (0.937 +/- 0.136 g/cm2, P-value 0.010; 0.875 +/- 0.113 g/cm2, P-value 0.045, respectively) than did women without MS. These differences at the spine and femoral neck were persisted after adjusting for age, body mass index, menopause status, alcohol consumption, smoking and physical activity. However, no significant differences in BMD were found in men between those with and without MS. In linear regression analyses, waist circumferences were highly associated with BMD at lumbar and total hip sites in women (beta -0.003, P-value < 0.001; beta -0.001, P-value 0.001, respectively). However, no significant differences were found in men between BMD and components of the MS. CONCLUSION: Among components of MS, waist circumference was associated with BMD in women. But no association was found in men. Women with MS had lower BMD than did women without MS.

The effect of pomegranate on postmenopausal syndrome in ovariectomized rats / 대한산부인과학회지

OBJECTIVE: To determine whether or not pomegranate extract can affect postmenopausal syndrome in ovariectomized rats. METHODS: Seventy-nine female Sprague-Dawley rats were used: A, no intervention; B, sham operation and distilled water; C, ovariectomy and distilled water; D, ovariectomy and 10% dilute pomegranate; E, ovariectomy and 20% pomegranate; and F, ovariectomy and 40% pomegranate. The study samples were obtained 4 weeks later. The following parameters were investigated for analyses: the thickness of urogenital epithelium, tail skin temperature, serum levels of sex hormones, lipid profile, homocysteine, and thiobarbituric acid reactive substances. RESULTS: 20% pomegranate had a transient thickening effect of bladder epithelial layer. There was an intermittent lowering effect of tail skin temperature of pomegranate extract, however, it was not consistent. There were lowering effect in serum homocysteine and elevating effect in serum high density lipoprotein-cholesterol. Pomegranate did not reverse post-ovariectomy reduced sex hormone levels. CONCLUSION: Pomegranate extract showed a tendency to reverse ovariectomy-related urothelial changes. Also pomegranate had a serum homocysteine-lowering effect.

The Effect of Pomegranate on Postmenopausal Syndrome: A Randomized, Double-blind, Placebo-controlled Trial / 대한폐경학회지

OBJECTIVES: To investigate the safety and effect of pomegranate extract on postmenopausal syndrome. METHODS: One hundred twelve women participated in a randomized, double-blind, placebo-controlled trial conducted in a tertiary university hospital in Korea. Women were treated with pomegranate extract or placebo for 12 weeks. The primary end point was menopausal symptoms, which were evaluated based on the Kupperman Index. The secondary end point was the serum estradiol (E2, pg/ml) level. Statistical analysis was performed. RESULTS: Menopausal symptoms as the Kupperman Index were significantly improved in the pomegranate group compared to placebo during the 12 weeks of intervention (P < 0.0001). Melancholia did not differ between the two groups. There was also no difference in serum E2 levels. There were no differences between the two groups in age, body weight, height, systolic and diastolic blood pressures, serum white and red blood cell counts, and fasting blood glucose, hemoglobin, aspartic transaminase, alanine transaminase, total cholesterol, triglycerides, high- and low-density lipoprotein-cholesterol, creatinine, and E2 levels. The side effects of pomegranate extract were negligible. CONCLUSION: Pomegranate extract is safe and effective in the treatment of menopausal symptoms. Further studies on whether or not the beneficial effect of pomegranate extract is due to the estrogen component are needed.

The Effect of Pomegranate on Postmenopausal Syndrome: A Randomized, Double-blind, Placebo-controlled Trial / 대한폐경학회지

OBJECTIVES: To investigate the safety and effect of pomegranate extract on postmenopausal syndrome. METHODS: One hundred twelve women participated in a randomized, double-blind, placebo-controlled trial conducted in a tertiary university hospital in Korea. Women were treated with pomegranate extract or placebo for 12 weeks. The primary end point was menopausal symptoms, which were evaluated based on the Kupperman Index. The secondary end point was the serum estradiol (E2, pg/ml) level. Statistical analysis was performed. RESULTS: Menopausal symptoms as the Kupperman Index were significantly improved in the pomegranate group compared to placebo during the 12 weeks of intervention (P < 0.0001). Melancholia did not differ between the two groups. There was also no difference in serum E2 levels. There were no differences between the two groups in age, body weight, height, systolic and diastolic blood pressures, serum white and red blood cell counts, and fasting blood glucose, hemoglobin, aspartic transaminase, alanine transaminase, total cholesterol, triglycerides, high- and low-density lipoprotein-cholesterol, creatinine, and E2 levels. The side effects of pomegranate extract were negligible. CONCLUSION: Pomegranate extract is safe and effective in the treatment of menopausal symptoms. Further studies on whether or not the beneficial effect of pomegranate extract is due to the estrogen component are needed.

Prenatally Detected Congenital Perineal Mass Using 3D Ultrasound which was Diagnosed as Lipoblastoma Combined with Anorectal Malformation: Case Report

We report a case of prenatally diagnosed congenital perineal mass which was combined with anorectal malformation. The mass was successfully treated with posterior sagittal anorectoplasty postnatally. On ultrasound examination at a gestational age of 23 weeks the fetal perineal mass were found on the right side. Any other defects were not visible on ultrasonography during whole gestation. Amniocentesis was performed to evaluate the fetal karyotyping and acetylcholinesterase which were also normal. As the fetus grew up, the mass size was slowly increased more and more. At birth, a female neonate had a perineal mass on the right side as expected. During operation, the anal sphincteric displacement was found near the mass and reconstructed through posterior sagittal incision. This is the first reported case of prenatally diagnosed congenital perineal mass, after birth which was diagnosed as lipoblastoma and even combined with anorectal malformation. This case shows that it can be of clinical importance to be aware of this rare fetal perineal mass in prenatal diagnosis and counseling.

Relationship between Metabolic Syndrome and Bone Mineral Density in Middle-aged Women / 대한폐경학회지

OBJECTIVES: To determine the relationship between metabolic syndrome (MS) and bone mineral density (BMD) in middle-aged women. METHODS: Three hundred eight premenopausal women were included in this study, including 268 women without MS and 40 women confirmed to have MS according to the National Cholesterol Education Program-Adult Treatment Panel III (NCEP-ATP III) criteria. Clinical markers including anthropometric parameters, the serum lipid profile, and glucose level were determined and analyzed with the BMD at the lumbar spine and femoral neck. RESULTS: There was no significant difference in femoral neck or lumbar BMD between the controls and women with MS. In correlation tests, the body mass index (BMI) showed a significant association with the femoral (r = 0.209, P < 0.001) and lumbar BMD (r = 0.176, P = 0.002). In addition, the waist circumference (WC) was also significantly correlated with the femoral (r = 0.142, P = 0.012) and lumbar BMD (r = 0.113, P = 0.048). CONCLUSION: The BMI and WC showed a significant positive association with the femoral and lumbar spine BMD in premenopausal women. Further studies with a large population will be needed to elucidate the combined effect of MS on BMD in middle-aged women.

Stemness Evaluation of Mesenchymal Stem Cells from Placentas According to Developmental Stage: Comparison to Those from Adult Bone Marrow

This study was done to evaluate the stemness of human mesenchymal stem cells (hMSCs) derived from placenta according to the development stage and to compare the results to those from adult bone marrow (BM). Based on the source of hMSCs, three groups were defined: group I included term placentas, group II included first-trimester placentas, and group III included adult BM samples. The stemness was evaluated by the proliferation capacity, immunophenotypic expression, mesoderm differentiation, expression of pluripotency markers including telomerase activity. The cumulative population doubling, indicating the proliferation capacity, was significantly higher in group II (P<0.001, 31.7+/-5.8 vs. 15.7+/-6.2 with group I, 9.2+/-4.9 with group III). The pattern of immunophenotypic expression and mesoderm differentiation into adipocytes and osteocytes were similar in all three groups. The expression of pluripotency markers including ALP, SSEA-4, TRA-1-60, TRA-1-81, Oct-4, and telomerase were strongly positive in group II, but very faint positive in the other groups. In conclusions, hMSCs from placentas have different characteristics according to their developmental stage and express mesenchymal stemness potentials similar to those from adult human BMs.

The Effect of Pomegranate on Depression, Anxiety, and Food Intake in Ovariectomized Rats / 대한폐경학회지

OBJECTIVES: To determine whether or not pomegranate extract can affect depression, anxiety, and food intake in ovariectomized rats. METHODS: Seventy-nine female Sprague-Dawley rats were divided into six groups: A, no operation and no drug intake; B, sham operation and distilled water; C, ovariectomy and distilled water; D, ovariectomy and 10% dilute pomegranate extract; E, ovariectomy and 20% dilute pomegranate extract; and F, ovariectomy and 40% dilute pomegranate extract. Beginning 2 days after surgery, drugs were administrated for 4 weeks. After that, the rats were subjected to the elevated plus maze (EPM) test and forced swim test (FST). RESULTS: The 10% pomegranate extract had a lower % closed arm entry frequency in the EPM test. A pomegranate dose-dependent decrease in the duration of immobility duration in FST was shown. Pomegranate did not reverse ovariectomy-related hyperphagia and weight gain. CONCLUSION: Pomegranate extract improved depression and anxiety in a postmenopausal model with ovariectomized rats.

Clinical Implication of 2nd Trimester Glycosuria / 대한주산의학회잡지

OBJECTIVE: To analyze the incidence of gestational diabetes mellitus (GDM) and its clinical implication of glycosuria identified in 2nd trimester pregnancy. METHODS: This study included pregnant women who had undertaken the 50 g oral glucose tolerance test (50 g OGTT) between 24 and 28 weeks gestation and delivered at term (N=704). Blood and urine sample were collected and analyzed for glucose level, one hour after 50 g OGTT. We applied women to 100 g OGTT if their blood glucose level after 50 g OGTT were more than 140 mg/dL. We compared blood glucose level, rate of GDM, birth weight and number of macrosomia at different urine glucose levels. Urine glucose level were measured by urine dipstick test and grouped to trace, 1+, 2+, 3+, and 4+, which were corresponding to 100, 250, 500, 1,000, 2,000 mg/dL. RESULTS: Women with glycosuria after 50 g OGTT were 258/704 (36.6%). Mean blood glucose levels were 117+/-23 mg/dL, 128+/-20 mg/dL, 135+/-23 mg/dL, 132+/-17 mg/dL, 139+/-25 mg/dL, 153+/-45 mg/dL, mean birth weight 3.29+/-0.40 kg, 3.25+/-0.40 kg, 3.27+/-0.41 kg, 3.34+/-0.35 kg, 3.28+/-0.41 kg, 3.33+/-0.40 kg, and numbers of macrosomia (> or =4.0 kg) 20 (4.5%), 3 (4.8%), 1 (1.8%), 2 (4.2%), 3 (6.7%), 0 (0%) at glycosuria level of negative, trace, 1+, 2+, 3+ and 4+ respectively. Glycosuria level was correlated significantly with blood glucose level (P=0.000), but not with birth weight and macrosomia (P=0.838, 0.881). The rate of GDM was 7/55 (12.7%), 2/48 (4.7%), 7/45 (15.6%), 8/48 (16.7%) in glycosuria level of 1+, 2+, 3+, 4+ and their relationship was statistically significant (P=0.000, AUC=0.734, 95%CI 0.638-0.830). In the cut off value of glycosuria 1+ or greater, sensitivity and positive predictive value were 72.7 and 12.2%. CONCLUSION: Glycosuria correlates well with blood glucose level and GDM prevalence but not with birth weight.

Adenocarcinoma arising in mature cystic teratoma: a case report / 부인종양

Benign cystic teratoma is recognized as one of the most common tumors in women during the reproductive age and frequently is treated by pelviscopic operation. Malignant transformation of a benign cystic teratoma is a rare event, and adenocarcinoma is extremely rare, and distinguishing this malignant change from benign disease preoperatively is nearly impossible even by the use of radiological imaging or various tumor markers. Therefore, patients should be informed that if a laparoscopic cystectomy is undertaken, a prompt second staging operation should be performed if the definitive pathology reveals an unexpected malignancy. We present a case with thyroid papillary carcinoma of follicular variant arising from mature cystic teratoma removed by laparoscopic salpingo-oophorectomy followed by staging laparotomy. We briefly reviewed literatures with regard to malignant transformation of a benign cystic teratoma.

Diagnosis and treatment of Pelvic Organ Prolapse / 대한산부인과학회지

Pelvic organ prolapse is defined anatomically as the descent of a pelvic organ into or beyond the vagina, perineum or anal canal. As a society is industrialized and the average span of life is increased, there are a growing numbers and concerns with pelvic organ prolapse. It can endanger our quality of life not only by a specific mechanical symptom, such as bulge outside the vagina, but by functional symptoms like bladder, bowel and sexual problems. Growth in demand for services to care for pelvic organ prolapse generates a demand for objective diagnostic tool and successful treatment strategies. So, first, we would like to discuss the POP Q system which is by far the most often used by clinicians and researchers since it's adoption in 1996, and second, we would like to compare the types, characteristics, success rates, and recurrence rates of two broad categories of prolapse management, nonsurgical treatment, which includes pelvic floor muscle training and pessary use, and surgery for anterior, apical and posterior vaginal wall defects.

The effectiveness of ethylene glycol as cryoprotectant in mouse embryo freezing with slow freezing method / 대한산부인과학회지

OBJECTIVE: We intended to know how the cryoprotectant ethylene glycol (EG) would affect the outcome of the embryo development when used in slow freezing method. And to know if there is any difference in the outcome of frozen-thawed embryos according to freezing methods and the timing. METHODS: We used 5-6 weeks old ICR female mice and T6 containing 0.4% BSA for basic culture media. The embryos at the developmental stages of 1-cell, 8-cell and blastocyst were cryopreserved respectively by slow freezing method using EG, propylene glycol (PROH), and glycerol as a cryoprotectant. We also compared the results of slow freezing and vitrification methods with the same cryoprotectant, EG. And finally, we evaluated the quality of blastocysts by counting the cell numbers in each group. RESULTS: The post-thaw embryo development were better in EG group when they were frozen at 1-cell and blastocyst stage (P<0.05). Although there were no differences in the recovery rate, the survival rate in vitrification group was significantly higher (P<0.05). Post-thaw embryo development to morula and blastocyst were better in vitrification group when frozen at 1-cell embryo (P<0.05), not at 8-cell and blastocyst group. The cell counts of blastocyst derived from 1-cell stage frozen EG group were significantly increased than that of PROH-glycerol groups (P<0.05), however, there was no difference between the two freezing methods. CONCLUSION: These results suggest that EG may be advantageous comparing with the conventional cryoprotectants, PROH and glycerol in slow freezing method for mouse embryo cryopreservation. In terms of freezing method, vitrification is better than slow freezing.

The study of the reproductive outcome after myomectomy in infertile patients with uterine myoma / 대한산부인과학회지

OBJECTIVE: The purpose of this study is to evaluate the reproductive outcome after myomectomy in infertile patients with uterine myoma. METHODS: During the period from January 1993 to December 2002, 55 infertile women with uterine myoma underwent abdominal myomectomy at Department of Obstetrics and Gynecology. RESULTS: Pregnancy occurred in 32 women and pregnancy rate was 58.2%. Parity, the number, type, and location of myoma, the opening of endometrial cavity during operation and the presence of endometriosis were not significantly different between the pregnant and nonpregnant group. But the mean age and duration of infertility were significantly low in pregnant group. The 24-month cumulative probability of conception was 84.4% and the spontaneous abortion rate after myomectomy was 28.1%. CONCLUSION: Our results suggest a benefit of myomectomy in infertile patients. Factors affecting the pregnancy rate after surgery in these patients are the age of the patient and the duration of infertility. Therefore, it is important that we should explain these factors to patients carefully before surgery.

Abnormal Uterine Bleeding

The average length of menstrual cycle is 28 days, and duration of flow is 4 days, and an average blood loss is 35cc. Abnormal uterine bleeding is defined as bleeding at abnormal or unexpected times or by an excessive flow at the time of expected menses. Any bleeding should be considered abnormal in premenarchal girls and in postmenopausal women. The etiology of abnormal uterine bleeding encompasses a wide range of disorders that can be secondary to pregnancy-related disorders, anatomic changes of the female genital tract, infection, endocrinologic disorders, malignancies, and systemic illnesses. Dysfunctional uterine bleeding (anovulatory or ovulatory) is diagnosed by exclusion of these causes. An appropriate workup is guided by age-related differential diagnoses for abnormal bleeding. Modern diagnostic tools can readily reveal the underlying pathology and allow timely intervention. Most abnormal genital tract bleeding occurs in the form of uterine bleeding, which is one of the most common gynecologic problems that health care providers encounter with, accounting for approximately 19% of office visits and 25% of gynecologic operations. The author will review the categories of abnormal uterine bleeding and the diagnostic tools needed to establish the correct diagnosis and treatment strategy.

Usefulness of an Unconjugated Estriol for prediction of Gestational Diabetes Mellitus / 대한산부인과학회지

OBJECTIVE: Gestational diabetes mellitus (GDM) is one of the most common metabolic disorders complicating pregnancy. It is associated with adverse outcomes of pregnancy including obstetrical complications such as increased rate of cesarean section, preeclampsia, and birth trauma, and perinatal morbidities, such as macrosomia, hypoglycemia, hypocalcemia, and hyperbilirubinemia. Therefore, screening for gestational diabetes mellitus and early diagnosis of this condition allows intervention to be carried out, thereby, the reduction of the untoward effects mentioned above can be minimized. But selective screening based on clinical or historic risk factors has been reported ineffective to identify the women with GDM. BACKGROUND: Circulating sex hormones have a role in the development of insulin resistance associated with certain physiological states including hyperandrogenism and polycystic ovary syndrome. Throughout pregnancy, normal human pregnancy is a hyperestrogenic state of major proportions. To evaluate the clinical correlation between Gestational diabetes mellitus and unconjugated estriol, we used it to screening protocol using three biochemical markers for unconjugated estriol levels. METHODS: In our retrospective between January 2002 and December 2003 at Korea university medical center, 137 women were screened for three biochemical tests and 50 gm Glucose challenge test at second trimester of pregnancy. The 50 gm GCT positive (n=56) women were performed 100 gm oral glucose tolerance test and by NDDG diagnosic criteria, women were diagnosed as Gestational diabetes mellitus (n=42). RESULTS: With logistic regression analysis, the significant risk factors were family history of DM (odd ration 16.59 95% CI 2.66-103.52), previous macrosomia birth (odd ratio 9.02 95% CI 1.98-41.6), maternal BMI (odd ratio 1.29 95% CI 1.09-1.49), parity (odd ratio 0.31 95% CI 0.11-0.83), glucosuria (odd ratio 0.68 95% CI 0.26-1.76) and among three biochemical test, estriol (odd ratio 1.60 95% CI 0.49-5.27), AFP (odd ratio 1.35 95% CI 0.35-5.28), hCG (odd ratio 0.59 95% CI 0.27-1.28). CONCLUSION: The screening test of GDM would be preferable and intensified management approach needed for patients with obesity, previous macrosomia birth. In three biochemical test, estriol and AFP levels were elevated in GDM but these odd ratio were not statistically significant.

Hematopoietic Differentiation of Embryoid Bodies Derived from the Human Embryonic Stem Cell Line SNUhES3 in Co-culture with Human Bone Marrow Stromal Cells

Human embryonic stem (ES) cells can be induced to differentiate into hematopoietic precursor cells via two methods: the formation of embryoid bodies (EBs) and co-culture with mouse bone marrow (BM) stromal cells. In this study, the above two methods have been combined by co-culture of human ES-cell-derived EBs with human BM stromal cells. The efficacy of this method was compared with that using EB formation alone. The undifferentiated human ES cell line SNUhES3 was allowed to form EBs for two days, then EBs were induced to differentiate in the presence of a different serum concentration (EB and EB/high FBS group), or co- cultured with human BM stromal cells (EB/BM co-culture group). Flow cytometry and hematopoietic colony-forming assays were used to assess hematopoietic differentiation in the three groups. While no significant increase of CD34+/CD45- or CD34+/CD38- cells was noted in the three groups on days 3 and 5, the percentage of CD34+/CD45- cells and CD34+/ CD38- cells was significantly higher in the EB/BM co-culture group than in the EB and EB/high FBS groups on day 10. The number of colony-forming cells (CFCs) was increased in the EB/BM co-culture group on days 7 and 10, implying a possible role for human BM stromal cells in supporting hematopoietic differentiation from human ES cell-derived EBs. These results demonstrate that co-culture of human ES-cell-derived EBs with human BM stromal cells might lead to more efficient hematopoietic differentiation from human ES cells cultured alone. Further study is warranted to evaluate the underlying mechanism.