ABSTRACT
@#【Objective】 A human embryonic stem cell line derived from Preimplantation genetic testing （PGT） embryos was established in a xeno- free stem cell culture system to provide disease models for medical research. 【Methods】The xeno-free culture system using xeno-free human foreskin fibroblast feeder layers（XF-HFF）mixed with commercially available chemically-defined medium（CDM）was assessed. In the culture system，a new hESC cell line was established using discarded embryos derived from PGT in patients with chromosomal balance translocation.【Results】The new availabled stem cell line was successfully cultured in the xeno-free culture system for a long time（> 45 passages）. The karyotype analysis revealed that the new line kept the same karyotype over 45 passages. Moreover，the expression of pluripotent markers was detected by fluorescent immunostaining including SSEA- 3，SSEA- 4，SSEA- 1，TRA- 1- 60， and TRA-1-81. RT-PCR analysis showed that the stem cell markers were present in hESC grown on XF-HFF-CDM. In addition，the teratoma formation analysis demonstrated that the cells cultured in XF-HFF/CDM maintained their pluripotency in vivo.【Conclusions】Our study may provide the possibility to establish embryonic stem cells with certain pathogenic genes，which could be applied for clinical research and treatment.
ABSTRACT
OBJECTIVE: To compare the clinical pregnancy rates between two types of endometrial preparation protocolsnatural cycle（NC）and hormone replacement cycle（HRT）-in patients with thin endometrium in the frozen-thawed embryo transfer（FET）cycles.METHODS: From January 2012 to December 2018,FET patients with endometrial thickness ≤7 mm on the day of human chorionic gonadotropin（h CG）trigger in Reproductive Medicine Center of the First Affiliated Hospital of Sun Yat-sen University were selected as research subjects.According to the endometrial preparation protocols,they were divided into NC group and HRT group.Totally 117 pairs were successfully matched using the propensity score matching method.The matching variables were age,embryo type and number of transferred embryos,and the embryo implantation rate and clinical pregnancy rate of the two matched groups were compared.RESULTS: There was no significant difference in embryo implantation rate（36.47% vs. 39.03%）or clinical pregnancy rate（44.40% vs. 52.10%）between NC group and HRT group（P> 0.05）.CONCLUSION: NC group and HRT group had similar pregnancy rate in patients with thin endometrium in FET cycles.Individualized protocols can be adopted according to the characteristics of patients with thin endometrium.
ABSTRACT
<p><b>BACKGROUND</b>Human embryonic stem (HES) cell derived from human blastocyst can be propagated indefinitely in the primitive undifferentiated state while remaining pluripotent. It has exciting potential in human developmental biology, drug discovery, and transplantation medicine. But there are insufficient HES cell lines for further study.</p><p><b>METHODS</b>Three oocyte donors were studied, and 3 in vitro fertilization (IVF) cycles were carried out to get blastocysts for the establishment of HES cell line. Isolated from blastocysts immunosurgically, inner cell mass (ICM) was cultured and propagated on mouse embryonic fibroblasts (MEFs). Once established, morphology, cell surface markers, karyotype and differentiating ability of the cell line were thoroughly analyzed.</p><p><b>RESULTS</b>Four ICMs from 7 blastocysts were cultured on MEFs. After culture, one cell line (cHES-1) was established and met the criteria for defining human pluripotent stem cells including a series of markers used to identify pluripotent stem cells, morphological similarity to primate embryonic stem cells and HES reported else where. Normal and stable karyotype maintained over 60 passages, and demonstrated ability to differentiate into a wide variety of cell types.</p><p><b>CONCLUSIONS</b>HES cell lines can be established from gamete donors at a relatively highly efficient rate. The establishment will exert a widespread impact on biomedical research.</p>
Subject(s)
Female , Humans , Male , Blastocyst , Cell Biology , Cell Differentiation , Cell Line , DNA-Binding Proteins , Fertilization in Vitro , Karyotyping , Octamer Transcription Factor-3 , Stem Cells , Cell Biology , Tissue Donors , Transcription FactorsABSTRACT
<p><b>BACKGROUND</b>The efficiency of traditional cryopreservation of human embryonic stem (ES) cells is low, and there have been few attempts to prove new cryopreservation methods effective. This study was designed to evaluate the efficiency of cryopreservation of human ES cells using vitrification method.</p><p><b>METHODS</b>Human ES cells clumped from an identical cell line were randomly allocated to be cryopreserved by vitrification or by slow freezing. The recovery rates, the growth and differentiation potential of thawed human ES cells were compared between these two groups. The pluripotency of human ES cells after thawing was identified.</p><p><b>RESULTS</b>Eighty-one point nine percent (59/72) of human ES cell clumps were recovered after vitrification, while only 22.8% (16/70) were recovered after slow freezing (P < 0.01). The colonies after vitrification manifested have not only faster growth but also a lower level of differentiation when compared to colonies subjected to the slow freezing protocol. However, the rates of growth and differentiation in undifferentiated colonies from both groups were identical to the rates in those of non-cryopreserved stem cells after a prolonged culture period. Passage 6 of vitrified human ES cells retained the properties of pluripotent cells, a normal karyotype and expressed the transcription factor OCT-4, stage specific expressed antigen-4 (SSEA-4) and SSEA-3. Teratoma growth of these cells demonstrated the ability to develop into all three germ layers.</p><p><b>CONCLUSIONS</b>Vitrification is effective in cryopreserving human ES cells. During a prolonged culture, human ES cells retain their pluripotency after cryopreservation.</p>