Transplantation of human placental chorionic plate-derived mesenchymal stem cells for repair of neurological damage in neonatal hypoxic-ischemic encephalopathy

Neonatal hypoxic-ischemic encephalopathy is often associated with permanent cerebral palsy,neurosensory impairments,and cognitive deficits,and there is no effective treatment for complications related to hypoxic-ischemic encephalopathy.The therapeutic potential of human placental chorionic plate-derived mesenchymal stem cells for various diseases has been explored.However,the potential use of human placental chorionic plate-derived mesenchymal stem cells for the treatment of neonatal hypoxic-ischemic encephalopathy has not yet been investigated.In this study,we injected human placental chorionic plate-derived mesenchymal stem cells into the lateral ventricle of a neonatal hypoxic-ischemic encephalopathy rat model and observed significant improvements in both cognitive and motor function.Protein chip analysis showed that interleukin-3 expression was significantly elevated in neonatal hypoxic-ischemic encephalopathy model rats.Following transplantation of human placental chorionic plate-derived mesenchymal stem cells,interleukin-3 expression was downregulated.To further investigate the role of interleukin-3 in neonatal hypoxic-ischemic encephalopathy,we established an in vitro SH-SY5Y cell model of hypoxic-ischemic injury through oxygen-glucose deprivation and silenced interleukin-3 expression using small interfering RNA.We found that the activity and proliferation of SH-SY5Y cells subjected to oxygen-glucose deprivation were further suppressed by interleukin-3 knockdown.Furthermore,interleukin-3 knockout exacerbated neuronal damage and cognitive and motor function impairment in rat models of hypoxic-ischemic encephalopathy.The findings suggest that transplantation of hpcMSCs ameliorated behavioral impairments in a rat model of hypoxic-ischemic encephalopathy,and this effect was mediated by interleukin-3-dependent neurological function.

Uterine epithelioid trophoblastic tumor with the main manifestation of increased human chorionic gonadotropin:A case report

BACKGROUND Epithelioid trophoblastic tumor(ETT)is an extremely rare malignant gestational trophoblastic neoplasm commonly presenting with abnormal vaginal bleeding,abdominal pain,and increased human chorionic gonadotropin(hCG).This study reported a case of uterine ETT with the main manifestation being increased hCG.CASE SUMMARY A 39-year-old female was referred to the Ningbo Maternal and Child Hospital of China in December 2022,complaining of increased hCG levels for 1 month.Magnetic resonance imaging revealed gestational trophoblastic tumor,and hysteroscopic electrotomy and curettage of intrauterine hyperplasia were performed.The patient was diagnosed with uterine ETT through postoperative pathological examination and immunohistochemical results.Total laparoscopic hysterectomy and bilateral salpingectomy were performed,and hCG levels returned to normal.The patient was without recurrence during the postoperative 3-month follow-up.CONCLUSION This study reported a case of uterine ETT with the main manifestation being increased hCG,highlighting that ETT should be considered in the presence of abnormal hCG.A total laparoscopic hysterectomy is recommended.

Predictive Value of Initial Serum Human Chorionic Gonadotropin Levels for Pregnancies after Single Fresh and Frozen Blastocyst Transfer

As one of the earliest markers for predicting pregnancy outcomes, human chorionic gonadotropin(h CG) values have been inconclusive on reliability of the prediction after frozen and fresh embryo transfer(ET). In this retrospective study, patients with positive h CG(day 12 after transfer) were included to examine the h CG levels and their predictive value for pregnancy outcomes following 214 fresh and 1513 vitrified-warmed single-blastocyst transfer cycles. For patients who got clinical pregnancy, the mean initial h CG value was significantly higher after frozen cycles than fresh cycles, and the similar result was demonstrated for patients with live births(LB). The difference in h CG value existed even after adjusting for the potential covariates. The area under curves(AUC) and threshold values calculated by receiver operator characteristic curves were 0.944 and 213.05 m IU/m L for clinical pregnancy after fresh ET, 0.894 and 399.50 m IU/m L for clinical pregnancy after frozen ET, 0.812 and 222.86 m IU/m L for LB after fresh ET, and 0.808 and 410.80 m IU/mL for LB after frozen ET with acceptable sensitivity and specificity, respectively. In conclusion, single frozen blastocyst transfer leads to higher initial h CG values than single fresh blastocyst transfer, and the initial h CG level is a reliable predictive factor for predicting IVF outcomes.

Effect of Mifepristone on the Telomerase Activity in Chorion and Decidua during Early Pregnancy

Objective To investigate telomerase activity in chorion and decidua from abortion induced by mifepristone incorporated with misoprostol at early pregnancy. Methods TRAP-SYBR Green assay was used to detect the expression of telomerase. Forty specimen were obtained from medicinal abortion （experiment group） and forty were from normal induced abortion （control group）. Results Positive expression of chorion telomerase was significantly different between the experimental group （28%, 11/40） and the control group （73%, 29/40） （P〈0.05）. While in decidua, the positive rate was 28% （11/40） in the experimental group and 20% （9/40） in the control group, there was no significant difference （P〉0.05）. Conclusion It is suggested that miferistone may significantly decrease the telomerase activity in chorion but not in decidua.

Second-trimester maternal β-human chorionic gonadotropin level associated with subsequent development of pregnancy-induced hypertension

Objective:To determine whether maternal β-human chorionic gonadotropin(β-hCG) level in second-trimester may be associated with subsequent development of pregnancy-induced hypertension(PIH).Methods:Seven hundred and sixty-two women in mid-trimester were to have maternal urine β-hCG standardized concentrations and maternal serum β-hCG measurements.Their case histories were recorded and reviewed from mid-trimester to delivery.The relation was observed between maternal urine,serum markers and subsequent development of PIH.Results:Among 762 women,504 cases were normal pregnancies,42 cases had PIH,94 cases had premature rupture of membrane (PROM),69 cases had preterm delivery (PD),53 other cases were excluded by various reasons.The levels of maternal urine,serum β-hCG in PIH were (61.75±9.78) IU/L and (304.56±54.17) ng/mg respectively,which were higher significantly than normal pregnancy group ([20.65±7.61] IU/L and [146.34±47.81] ng/mg,P<0.05).When maternal serum,urine β-hCG levels ≥2 MOM(multiple of mean),the incidences of developing PIH were increased significantly as compared with those of β-hCG <2 MOM women.The incidence of PIH increased from 5.1% in pregnancies with urine β-hCG ≥2 MOM to 11.7% in cases with urine β-hCG ≥4 MOM.Conclusion:The elevation of maternal mid-trimester urine,serum β-hCG levels is not only an early signal for dysfunction of placenta but also a dangerous signal for development of PIH.Second-trimester maternal urine β-hCG measurement proves to be superior to serum marker in clinical prediction.

Clonal isolation of endothelial colony-forming cells from early gestation chorionic villi of human placenta for fetal tissue regeneration

BACKGROUND Endothelial colony-forming cells(ECFCs)have been implicated in the process of vascularization,which includes vasculogenesis and angiogenesis.Vasculogenesis is a de novo formation of blood vessels,and is an essential physiological process that occurs during embryonic development and tissue regeneration.Angiogenesis is the growth of new capillaries from pre-existing blood vessels,which is observed both prenatally and postnatally.The placenta is an organ composed of a variety of fetal-derived cells,including ECFCs,and therefore has significant potential as a source of fetal ECFCs for tissue engineering.AIM To investigate the possibility of isolating clonal ECFCs from human early gestation chorionic villi(CV-ECFCs)of the placenta,and assess their potential for tissue engineering.METHODS The early gestation chorionic villus tissue was dissociated by enzyme digestion.Cells expressing CD31 were selected using magnetic-activated cell sorting,and plated in endothelial-specific growth medium.After 2-3 wks in culture,colonies displaying cobblestone-like morphology were manually picked using cloning cylinders.We characterized CV-ECFCs by flow cytometry,immunophenotyping,tube formation assay,and Dil-Ac-LDL uptake assay.Viral transduction of CVECFCs was performed using a Luciferase/tdTomato-containing lentiviral vector,and transduction efficiency was tested by fluorescent microscopy and flow cytometry.Compatibility of CV-ECFCs with a delivery vehicle was determined using an FDA approved,small intestinal submucosa extracellular matrix scaffold.RESULTS After four passages in 6-8 wks of culture,we obtained a total number of 1.8×107 CV-ECFCs using 100 mg of early gestational chorionic villus tissue.Immunophenotypic analyses by flow cytometry demonstrated that CV-ECFCs highly expressed the endothelial markers CD31,CD144,CD146,CD105,CD309,only partially expressed CD34,and did not express CD45 and CD90.CV-ECFCs were capable of acetylated low-density lipoprotein uptake and tube formation,similar to cord blood-derived ECFCs(CB-ECFCs).CV-ECFCs can be transduced with a Luciferase/tdTomato-containing lentiviral vector at a transduction efficiency of 85.1%.Seeding CV-ECFCs on a small intestinal submucosa extracellular matrix scaffold confirmed that CV-ECFCs were compatible with the biomaterial scaffold.CONCLUSION In summary,we established a magnetic sorting-assisted clonal isolation approach to derive CV-ECFCs.A substantial number of CV-ECFCs can be obtained within a short time frame,representing a promising novel source of ECFCs for fetal treatments.

Increased Apoptosis in Chorionic Trophoblasts of Human Fetal Membranes with Labor at Term

Objective: To determine the association of apoptosis in the layers of human fetal membranes distal to rupture site with labor at term. Study Design: Fetal membranes were collected from elective cesarean sections (n = 8) and spontaneous vaginal deliveries (n = 8) at term. The extent of apoptosis within fetal membrane layers was determined using terminal deoxynucleotidyl transferase deoxy-UTP-nick end labeling (TUNEL) assay and western blots for pro-apoptotic active caspase-3 and anti-apoptotic Bcl-2. Results: Apoptotic index in chorionic trophoblasts of membranes distal to rupture site obtained after vaginal delivery was 3-fold higher than those obtained from elective cesarean (11.57% ± 4.98% and 4.05% ± 2.4% respectively;p = 0.012). The choriodecidua layers after vaginal deliveries had higher expression of the pro-apoptotic active caspase-3 and less expression of the anti-apoptotic Bcl-2 than those obtained from elective cesarean sections. Conclusions: Labor at term is associated with increased apoptosis in chorionic trophoblast cells of human fetal membranes distal to rupture site.

How and Why Do Gestational Trophoblastic Neoplasms Overproduce Human Chorionic Gonadotropin?

From the published data, the present mini-review attempts to answer two fundamental questions about the gestational trophoblastic neoplasms. In addition, it extrapolates the findings to other cancers that produce small amounts of hCG and how a novel therapies could be developed.

DETECTION OF STRAND BREAKS OF DNA IN HUMAN EARLY CHORIONIC VILLUS CELLS INDUCED BY DIAGNOSTIC ULTRASOUND USING ^(32)P-LABELED ALU HYBRIDIZATION

Objective To explore if strand breaks of DNA in human early chorionic villus cells in uterus were induced by diagnostic ultrasound and to evaluate the method used for detection of single-stranded breaks and double-stranded breaks in human DNA. Methods 60 normal pregnant women aged 20-30, who underwent artificial abortion during 6-8 weeks of gestation, were randomly divided into 2 experimental groups: All 30 cases were exposed to diagnostic ultrasound in uterus for 10 minutes, and 24 hours later chorionic villi were extracted; the other 30 cases were taken as the control group. Single-stranded DNA and double-stranded DNA in villus cells in all cases were isolated by the alkaline unwinding combined with hydroxylapatite chromatography, and were quantitatively detected using 32 P-labeled Alu probe for dot-blotting hybridization. Results There was no significant difference in quantity and percentage in single-stranded DNA and double-stranded DNA between 2 groups (P>0.05). 32 P-Alu probe could only hybridize with human DNA, and could detect DNA isolated from as few as 2.5×10 3 chorionic villus cells and 0.45ng DNA in human leukocytes. Conclusion The results suggested that there were no DNA strand damages in human chorionic villus cells when the uterus was exposed to diagnostic ultrasound for 10 minutes. The method,^(32)P-Alu probe for dot-blotting hybridization, was even more specific, sensitive and accurate than conventional approaches.

Identification of chorion genes and RNA interference-mediated functional characterization of chorion-1 in Plutella xylostella

Choriogenesis is the last step of insect oogenesis,a process by which the chorion polypeptides are produced by the follicular cells and deposited on the surface of oocytes in order to provide a highly specialized protective barrier to the embryo.The essential features of chorion genes have yet to be clearly understood in the diamondback moth,Plutella xylostella,a worldwide Lepidoptera pest attacking cruciferous crops and wild plants.In this study,complete sequences for 15 putative chorion genes were identified,and grouped into A and B classes.Phylogenetic analysis revealed that both classes were highly conserved and within each,branches are also species-specific.Chorion genes from each class were located in pairs on scaffolds of the P.xylostella genome,some of which shared the common promoter regulatory region.All chorion genes were highly specifically expressed in the P.xylostella adult females,mostly in the ovary with full yolk,which is a crucial period to build the shells of the eggs.RNAi-based knockdown of chorion-1,which is located on the Px_scaffold 6 alone,although had no effect on yolk deposition,resulted in smaller eggs and sharply reduced hatchability.Additionally,inhibition of PxCho-1 expression caused a less dense arrangement of the columnar layers,reduced exochorion roughness and shorter microvilli.Our study provides the foundation for exploring molecular mechanisms of female reproduction in P.xylostella,and for making use of chorion genes as the potential genetic-based molecular target to better control this economically important pest.

Expression of Beta-Human Chorionic Gonadotropin Genes in Renal Cell Cancer and Benign Renal Disease Tissues

To study the expression of beta-human chorionic gonadotropin (βhCG) genes in renal cell carcinomas (RCC) and benign renal disease tissues, nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction endonuclease analysis were employed to detect the expression of βhCG genes in 44 cases of RCC tissues and 24 cases of benign renal disease tissues It was found that 52% RCC samples revealed positive for βhCG mRNA expression Positive rate in advanced stage and poorly differentiated RCC was higher, but there was no significant difference The positive rate of βhCG mRNA expression was 54% in 24 cases of benign renal tissues, including 3 cases out of 6 polycystic kidneys, 7 cases out of 13 renal atrophies, 2 cases out of 2 oncocytomas and 1 case out of 2 pyonephrotic kidneys β7 was most frequently transcribed subtype gene independent on the histology These findings suggested βhCG gene transcription is not only involved in RCC but also in benign renal diseases

Effects on Cell Viability and on Apoptosis in Tumoral(MCF-7)and in Normal(MCF10A)Epithelial Breast Cells after Human Chorionic Gonadotropin and Derivated-Angiotensin Peptides Treatments

Angiotensin-(1 - 7) [Ang-(1 - 7)] is an endogenous heptapeptide hormone of the renin-angiotensin system that has antiproliferative properties. The aim of this work was to evaluate the anti-proliferative and pro-apoptotic properties of Ang-(1 - 7) and of Ang-(1 - 7)-substituents 9-fluorenylmethyloxycarbonyl (Fmoc) e Ang II-derivatives containing the TOAC (2,2,6,6-tetramethylpiperidine-N-oxyl-4-amino-4-carboxylic acid) in normal (MCF10A) and in tumoral (MCF7) epithelial mammary cell lines. Both cell lines received an hCG and angiotensin peptides 24-hour treatment, in combination or alone followed by cell viability, apoptosis and cell cycle assays performed by flow cytometer (GUAVA). After hCG, Ang-(1 - 7), hCG + Ang-(1 - 7) and hCG + Ang-(1 - 7)-Fmoc treatments, MCF7 displayed cell viability decrease and mid-apoptosis increase. We also observed cell viability decrease in MCF10A after Ang-(1 - 7), Ang-(1 - 7) Fmoc and hCG + AngII Toac treatments. These cells had an increase in late apoptosis and necrosis after AngII Toac, hCG + Ang-(1 - 7) and hCG + Ang-(1 - 7)-Fmoc treatments. Regarding the cell cycle analysis, we did not observed any changes in cell cycle phases. In summary, cell viability was decreased and apoptosis (initial, mid and late) was increased after hCG and/or Ang-(1 - 7) peptides treatments. These results point out hCG and Ang-(1 - 7) as effective compounds to inhibit cell proliferation, since they decrease cell viability and increase apoptosis in both normal and in tumoral breast cells, being the effect more pronounced in the tumoral cell line. Our results support the idea of investigating more closely the putative use of these compounds as novel therapeutic agents for breast cancer.

The Influences of Mifepristone, Norethisterone and Tamoxifen on the Glycosphingolipids Compositions from Human Chorionic Tissue during Early Pregnancy

By using the Ladisch partitioning and microscale-analysis of HPTLC,the comparative quantitative and qualitative studies of gangliosides (Gg)and neutral glycosphingolipids(N-GSL)compositions from human chorionic villi tissues of normal early pregnant women and women treated with mifepristone, norethisterone(NET)and tamoxifen(TAM)were reported in this paper.The patterns of Gg and N-GSL in three treated groups were similar to those in normal pregnant group.The total values of Gg from the chorionic villi tissues reduced significantly in three treated groups(P<0.01).In all treated groups,the amounts of NeuNAC-Gal-Glc-cer(GM3),NeuNAC-NeuNAC-Gal-Glc-cer(GD3)and Gal-GalNAC-(NeuNAC)-Gal-Glc-cer(GM1)were decreased significantly compared with those in normal(P<0.01orP<0.05).In NET and TAM groups,Neu NAC-Gal-GalNAC-(NeuNAC-NeuNAC)-Gal-Glc-cer(GT1b) was markedly lower than that in normal(P<0.01).The total values of N-GSL extracted from the chorionic villi tissues were obviously higher in mifepristone and TAM groups than those in normal(P<0.01).The Gal-Glc-cer(LacCer)(CDH)and Gal-Gal-Glc-cer(Gal-LacCer)(CTH)were greatly increased in mifepristone group as compared with normal P<0.05).Paragloboside:Gal-GalNAC-Gal-Glc-cer(PG)in NETgroup was significantly higher than that in normal(P<0.01).

The Expression of Integrin β_3 and Intercellular Adhesion Molecule(ICAM-1) in Decidua and Chorionic Villi during Mifepristone Induced Abortion

The effects of mifepristone with misoprostol on the expression of the integrin β 3 and intercellular adhesion molecule 1 (ICAM 1) in decidua and chorionic villi tissues in early pregnancy in 10 cases were investigated by immuno flow cytometry (the experiment group). At the same time, the other 10 cases induced by mechanical vacuum aspiration were collected as the control. The results showed that, the positive rate of integrin β 3 and ICAM 1 in decidua of the experiment group were 19.1±5.01% and 20.61 ±6.51%; while those in chorionic villi were 21.32±4.38% and 20.29± 6.49%, which were significantly lower than those in the control group. These results suggested that integrin β 3 and ICAM 1 may take part in the maintenance of early pregnancy. The mechanism of mifepristone induced abortion may be mediated by the down regulation of the integrin β 3 and ICAM 1 expression in decidua and chorionic villi.

Morphological Progression of Chorionic Membrane Development in Monochorionic and Dichorionic Twins: A Sonographic Assessment

Background: The zygote of twins implants themselves separately and on different spots in the uterine endometrium. However, the growth of fetal membranes may be collective or singly. There is little data from sonographic view of assessment on the exact progression at different stages of gestation. More so, the realistic evidences from radiographic assessment of the compartmentalization of chorion membrane in the developing monochorionic and dichorionic twins are not sufficiently available, hence, the call to ascertain the exact progression of the chorion membrane through ultrasound scanning in gestational subjects. Aim: This present study examined the structural progression of embryonic growth pattern of chorionic membrane in monochorionic and dichorionic twins. Materials and Methods: The study utilized transabdominal ultrasound to periodically assess the progression of chorion membrane and advancement in compartmentalization of monochorionic and dichorionic twins as pregnancy proceeds. Results: The monochorionic membrane showed an enclosure that progresses to be more distinct and thickened around the two embryos with a unique T-shaped point of insertion in latter development. The partitioning of dichorionic membrane progresses to be less thickened and with a distinct lambda (λ) sign which is a wedge-shaped protrusion into the inter-twin space. Conclusion: Assessment showed that the growth pattern of chorion membrane varied with different parameters observed as early as in the first trimester. Hence during early gestational stage, a twin can be said to be monochorionic or dichorionic with key anatomical landmarks monitored. The growth progression could be used to project abnormality and on time treatment would be offered to improve perinatal outcome.

Effect of different fertilization time after human chorionic gonadotropin injection on fertilization outcome of patients in vitro fertilizationembryo transfer

Objective:To explore the effect of different fertilization time after human chorionic gonadotropin(HCG)injection on the outcome of fertilization in vitro fertilization-embryo transfer(IVF-ET).Methods:One thousand one hundred and forty IVF-ET cycles from January 2016 to August 2018 were analyzed retrospectively.According to the different fertilization time after injection of HCG divided into four groups:Group A(38.0 h~39.0 h),Group B(39.1 h~40.0 h),Group C(40.1 h~41.0 h),and Group D(41.1 h~42.0 h).The normal fertilization rate,the normal cleavage rate,the embryo utilization rate,the high-quality embryo rate,the clinical pregnancy rate,the implantation rate,and the spontaneous abortion rate were analyzed among the groups.Then we investigated the effect of different promotion methods on the outcome of fertilization during the optimal fertilization time.Results:There was no significant difference in 2PN cleavage rate,available embryo rate,clinical pregnancy rate,implantation rate and abortion rate among the four groups(P>0.05).The high-quality embryo rate in Group D(44.6%)was the highest,and was significantly different among the four groups(P<0.05).The normal fertilization rate in Group D(71.6%)was the highest,and was significantly different among the four groups(P<0.05).The normal fertilization rate(78.1%)of antagonist group was significantly higher than other groups(P<0.05).Conclusion:The different fertilization time after HCG injection have effects on high-quality embryo rate and normal fertilization rate of patients in IVF-ET.The appropriate fertilization time of patients in IVF-ET was 41 h~42 h after HCG injection in our reproductive center,improved the clinical pregnancy rate and reduced the early abortion rate.The GnRH-ant protocol is superior to other protocol in IVF-ET.

Effect of TCDD on Maternal Toxicity and Chorionic Gonadotropin──Bioactivity in the Immediate Post-implantation Period of Macaque

The purpose of this experiment was to observe the alterations in bioactivity of chorionic gonadotropin (CG) associated with early fetal loss (EFL), induced by the environmental toxin TCDD (2,3,7,8\|tetrachlorodibenzo\| p \|dioxin) in the cynomolgus macaque. Ten of twelve females administered single doses of 1, 2 or 4 μg/kg TCDD on gestational day (GD) 12 had EFL from ten to twenty days later. Seven control animals treated only with the vehicle had normal pregnancies. Blood samples were repeatedly collected for hormone evaluation, from two days before treatment to thirty\|one days following treatment. Immunoreactive monkey chorionic gonadotropin (mCG) was measured in serum using ELISA, and bioactive mCG was measured using a luminescence LH/CG bioassay. No change in immunoreactive mCG levels was detected as a result of TCDD, treatment, but bioactive mCG levels were significantly lower in TCDD\|treated animals compared to controls. This change in bioactivity of mCG was also reflected in the ratio of mCG bioactivity to mCG immunoreactivity (B/I ratio) which began to rise in normal pregnancies by GD 20, but did not rise in TCDD treated animals. These results demonstrate that normal pregnancy in the monkey, as in humans, is characterized by a post\|implantation change in the B/I ratio of CG. These findings therefore suggest that changes in the production of bioactive CG may be used as a biomarker of environmental toxicant exposures which lead to EFL.

The Study on Immuno-response and Antisera Properties of Recombinant β-subunit of Human Chorionic Gonadotropin in C57 Black Mouse

In this study, the immuno-response of recombinant hCG-β and natural hCGβ was comparatively investigated by using Freund's adjuvant. The results showed that, the properties and merits of the antibodies elicited by both kinds of hCG-β were similar. The antisera had high affinity for binding with hCG (Kαγβ≈5.86×108/mol/L, Kαβ≈8.18×108/mol/L), and were found to be effective in inhibiting the binding of 125I-hCG to receptors in rat testes. Results also indicated that, similar to the antisera induced by natural hCG-β, the recombinant hCG-β induced antisera had capacity of neutralizing the biological activities of hCG. Recombinant hCG-β could be used as an immunogen for contraceptive vaccine.

Early gestation chorionic villi-derived stromal cells for fetal tissue engineering

AIM: To investigate the potential for early gestation placenta-derived mesenchymal stromal cells(PMSCs) for fetal tissue engineering.METHODS: PMSCs were isolated from early gestation chorionic villus tissue by explant culture. Chorionic villus sampling(CVS)-size tissue samples(mean = 35.93 mg)were used to test the feasibility of obtaining large cell numbers from CVS within a clinically relevant timeframe. We characterized PMSCs isolated from 6 donor placentas by flow cytometry immunophenotyping, multipotency assays, and through immunofluorescent staining. Protein secretion from PMSCs was examined using two cytokine array assays capable of probing for over 70 factors in total. Delivery vehicle compatibility of PMSCs was determined using three common scaffold systems: fibrin glue, collagen hydrogel, and biodegradable nanofibrous scaffolds made from a combination of polylactic acid(PLA) and poly(lactic-co-glycolic acid)(PLGA). Viral transduction of PMSCs was performed using a Luciferase-GFPcontaining lentiviral vector and efficiency of transduction was tested by fluorescent microscopy and flow cytometry analysis.RESULTS: We determined that an average of 2.09 × 106(SD ± 8.59 × 105) PMSCs could be obtained from CVS-size tissue samples within 30 d(mean = 27 d, SD ± 2.28), indicating that therapeutic numbers of cells can be rapidly expanded from very limited masses of tissue. Immunophenotyping by flow cytometry demonstrated that PMSCs were positive for MSC markers CD105, CD90, CD73, CD44, and CD29, and were negative for hematopoietic and endothelial markers CD45, CD34, and CD31. PMSCs displayed trilineage differentiation capability, and were found to express developmental transcription factors Sox10 and Sox17 as well as neuralrelated structural proteins NFM, Nestin, and S100 β. Cytokine arrays revealed a robust and extensive profile of PMSC-secreted cytokines and growth factors, and detected 34 factors with spot density values exceeding 103. Detected factors had widely diverse functions that include modulation of angiogenesis and immune response, cell chemotaxis, cell proliferation, blood vessel maturation and homeostasis, modulation of insulin-like growth factor activity, neuroprotection, extracellular matrix degradation and even blood coagulation. Importantly, PMSCs were also determined to be compatible with bothbiological and synthetic material-based delivery vehicles such as collagen and fibrin hydrogels, and biodegradable nanofiber scaffolds made from a combination of PLA and PLGA. Finally, we demonstrated that PMSCs can be efficiently transduced(> 95%) with a Luciferase-GFPcontaining lentiviral vector for future in vivo cell tracking after transplantation.CONCLUSION: Our findings indicate that PMSCs represent a unique source of cells that can be effectively utilized for in utero cell therapy and tissue engineering.

CHROMATOGRAPHIC SEPERATION AND PURIFICATION OF HUMAN CHORIONIC GONADOTROPIN

Seperation and purifitation of human chorionic gonadotropion (HCG) in theurine sample of early pregnant women by D3520 resin adsorption chromatographyis repaied. The crude product obtained by DEAE - Cellulose 23 and DEAE Sephadex A50 column chromatography showed a high activity of HCG. Furtherpurifikation of the sample by gel filtration chromatography on a Sephadex G75 column gives a final preparation of 6000～ 6500 IU/mg. The preparation meets therequiTements of the pyrogn test in Chinese Law of Pharmacopoeia.