Moving toward Xeno-free Culture of Human Pluripotent Stem Cells

Human pluripotent stem cells (hPSCs) were conventionally cultured on feeder cells that are isolated from mouse embryonic fibroblast (MEF). However, these culture components could contaminate the hPSCs and can limit the application of hPSCs for clinical use. On the other hand, we demonstrated that exogenous basic fibroblast growth factor (bFGF) could be omitted from the hPSC culture media if we used the suitable feeder cells. We also showed that although hPSCs can proliferate on the feeder-free culture system, however, genetic instability of hPSCs has been reported in such environment. Feeder cells enable hPSCs to maintain their pluripotency. The feeder cells are usually grown in a culture medium containing fetal bovine serum (FBS) prior to coculture with hPSCs. The use of FBS might limit the clinical application of hPSCs. We proposed the use of human cord blood-derived serum (hUCS) and showed a positive effect on culture of mesenchymal stem cells. The results showed that human foreskin fibroblasts (HFFs) cultured in hUCS-containing medium (HFF-hUCS) displayed fibroblastic features, short population doubling times, high proliferation rates, and normal karyotypes after prolonged culture. These studies of hPSC xeno-free culture have been growing in both basic research and clinical trial. The data regarding the current clinical trials of using hPSCs convince the researchers not only about the possibility of application of hPSCs for cell-based therapy, but also the quality of established hPSC lines. Most of the hPSC lines that were published in the literature and registered in the National Institute of Health (NIH), hPSCreg of the European Union are not Good Manufacturing Practice (GMP) grade cell lines. Since one of the goals of using hPSCs is therapeutic purpose, GMP for derivation, cultivation, and handling the hPSCs are required. This chapter also reviews the state-of-the-art xeno-free culture system of hPSCs in the respect of future clinical applications

KAP Study on Contraception in Teenage Pregnancy at Antenatal Care Clinic and Delivery Room in Chulalongkorn Memorial Hospital

Objective: To assess knowledge, attitude, and practice about contraception in pregnant teenagers.Materials and Methods: This was a cross-sectional descriptive study. Data collection started from September 2009 to March 2010. Pregnant teenagers, aged 12-19 years old, attending ANC clinic or delivery room at King Chulalongkorn Memorial Hospital were enrolled. The questionnaire consists of four parts; demographic data, contraceptive knowledge, attitudes about pregnancy and contraception, and past contraceptive practices. Data were analyzed using descriptive statistics, chi square test, t-test and Pearson correlation.Results: A total of 150 complete questionnaires from 285 cases of teenage pregnancy in this study period were collected and analyzed. The mean age of subjects was 17.7 years. Only 52.7% used any contraception at first intercourse. Contraceptive pill was the most common method used (59.5%) and only one-fourth chose condom. Up to 74% of participants were pregnant for the first time and 72% informed that their current pregnancies were unintended. The mean contraceptive knowledge score was 70.0 ± 9.7%. Their knowledge short-acting contraception was significantly more than long-acting contraception (P<.001). There was a correlation between knowledge and use of contraception at first intercourse (P=.012) and a trend of correlation between knowledge and attitude although not significant. Most of teenagers decided to use contraception by themselves. When they had any problems, only 19% went to see a doctor. Sources of contraceptive information were mainly from school teaching and mass media.Conclusion: Knowledge on contraception in pregnant teenagers was in average level. There was a correlation between knowledge and the use of contraception at first sexual intercourse. Most cases decided to use contraception themselves (63.5%). Pills was the most common contraceptive method (63.5%)

Knowledge and Attitude of Obstetricians to the Protection of Children Born from Assisted Reproductive Technology

Objectives: This study aims to examine the attitudes and knowledge of obstetricians in related to the Protection of Children Born from Assisted Reproductive Technologies Act 2015. Materials and Methods: This was a survey study using a mailed questionnaires sent to 2,550 Obstetricians who were registered with The Royal Thai College of Obstetricians and Gynaecologists (RTCOG) and sent to their registered addresses. The questionnaires consisted of knowledge and attitude questions.Results: Replies were received from 340 obstetricians with a response rate of 13.3%. Of these Thai Obstetricians, 81.5% had a good attitude to the law while 14.7% and 3.8% respectively had moderate and poor attitude. The majority (56.7%) had a good knowledge of the law, while 37.1% had a moderate knowledge and just 6.2% had a poor knowledge. Nearly one hundred percent strongly agreed that there should be a law to control ART, that the law will help monitor ART and maintain good morals as well as protecting the dignity of attending obstetricians. Conclusion: The responded obstetricians have a good attitude towards and knowledge of the Protection of Children Born from Assisted Reproductive Technologies Act 2015. The RTCOG should inform nationwide obstetricians about this law and provide data center support

Human Umbilical Cord Blood-Derived Serum for Culturing the Supportive Feeder Cells of Human Pluripotent Stem Cell Lines

Although human pluripotent stem cells (hPSCs) can proliferate robustly on the feeder-free culture system, genetic instability of hPSCs has been reported in such environment. Alternatively, feeder cells enable hPSCs to maintain their pluripotency. The feeder cells are usually grown in a culture medium containing fetal bovine serum (FBS) prior to coculture with hPSCs. The use of FBS might limit the clinical application of hPSCs. Recently, human cord blood-derived serum (hUCS) showed a positive effect on culture of mesenchymal stem cells. It is interesting to test whether hUCS can be used for culture of feeder cells of hPSCs. This study was aimed to replace FBS with hUCS for culturing the human foreskin fibroblasts (HFFs) prior to feeder cell preparation. The results showed that HFFs cultured in hUCS-containing medium (HFF-hUCS) displayed fibroblastic features, high proliferation rates, short population doubling times, and normal karyotypes after prolonged culture. Inactivated HFF-hUCS expressed important genes, including Activin A, FGF2, and TGFβ1, which have been implicated in the maintenance of hPSC pluripotency. Moreover, hPSC lines maintained pluripotency, differentiation capacities, and karyotypic stability after being cocultured for extended period with inactivated HFF-hUCS. Therefore, the results demonstrated the benefit of hUCS for hPSCs culture system

Immunostaining characterization and percentage of H5N1-positive cells of hNPCs following virus infection.

<p>Virus antigens were distributed within the cytoplasm and the nucleus of the infected cells (<b>A-C</b>), with peak percentage of virus-positive cells about 75% at 24 hpi, then declined to about 30% at 72 hpi (<b>D</b>). Scale bar in A-C ≈ 30 μm. Data represented are the mean ± standard error. Asterisks indicated statistically significant differences (<i>p</i>-value<0.05).</p