Objective To evaluate the effects of different sera on the growth of human granulosa cells （GCs） cultured in vitro. Methods GCs were obtained from women who underwent follicular aspirates during in vitro fertilizati...Objective To evaluate the effects of different sera on the growth of human granulosa cells （GCs） cultured in vitro. Methods GCs were obtained from women who underwent follicular aspirates during in vitro fertilization （IVF） program. Five groups were divided according to media supple- mented with different types of sera. Group A, 10% heat-inactivated fetal bovine serum （FBS）; group B, 10% heat-inactivated newborn bovine serum; group C, 10% non heat-inactivated FBS; group D, 10% human serum; group E, bovine serum albumin. Morphological characteristics and viability of GCs measured by trypan blue exclusion assay were evaluated after 24 h of incubation.Results GCs cultured in group A and group B showed multiform morphology compared with other groups. GCs cultured in group D and group E were present with cytoplasmic atrophy and less pseudopodium. Moreover, group A and group B showed a similar level in the viability of GCs （P〉0.05）, which displayed no difference between group D and group E as well （P〉0.05）. Group C had a lower level of viability than group A （P〈0.05） but a higher level than group D （P〈0.01）. Conclusion Heat-inactivated sera can improve the growth of GCs. Different types of sera would have different effects on the growth of GCs cultured in vitro. The pre-culture with different types of sera should be performed to get better efficacy.展开更多
Background： In the current society, infertility related to age has become a social problem. The in vitro fertilization （IVF） success rate in women with poor ovarian response （POR） is very low. Dandelion extract T...Background： In the current society, infertility related to age has become a social problem. The in vitro fertilization （IVF） success rate in women with poor ovarian response （POR） is very low. Dandelion extract T-1 （DE-T1） is an effective component of the extract from the leaves and stems of Traxacum officinale, which is one of the medicines used in some patients with POR, but its molecular mechanism remains unclear. Methods： Following IVF, ovarian granulosa cells （GCs） of sixty patients were extracted and divided into normal ovarian response （NOR） and POR groups. GCs were cultured in a dose-dependent and time-dependent manner with DE-TI, proliferation of GCs was determined by Cell Counting Kit-8 assay, and mRNA levels of insulin-like growth factor 1 receptor （IGF-1R）, luteotropic hormone receptor （LHR）, follicle-stimulating hormone receptor （FSHR）, LHR, and CYP19A1 （aromatase） were determined by quantitative polymerase chain reaction. Progesterone and estradiol （E2） concentrations were determined by enzyme-linked immunosorbent assay. Results： The cell viability gradually increased with the progressive increase in the DE-T1 concentration. Compared with the control group （without DE-T1）, the mRNA expressions of FSHR, LHR, IGF-IR, and CYPIgAI were upregulated after the addition of DE-T l, especially in the 2.5% DE-T 1 group （P 〈 0.01 ）. The expression of IGF- 1R was upregulated approximately 25 times （24.97 ± 4.02 times） in the POR group with 2.5% DE-T1. E2 and progesterone levels increased with the increasing DE-T1 concentration. There were highly significant differences in the E2 and progesterone secretion between the NOR and POR groups （P 〈 0.01）. Conclusion： DE-TI may promote steroid hormone synthesis by promoting GC proliferation and upregulating GC receptor expression, thereby improving ovarian endocrine function.展开更多
文摘Objective To evaluate the effects of different sera on the growth of human granulosa cells （GCs） cultured in vitro. Methods GCs were obtained from women who underwent follicular aspirates during in vitro fertilization （IVF） program. Five groups were divided according to media supple- mented with different types of sera. Group A, 10% heat-inactivated fetal bovine serum （FBS）; group B, 10% heat-inactivated newborn bovine serum; group C, 10% non heat-inactivated FBS; group D, 10% human serum; group E, bovine serum albumin. Morphological characteristics and viability of GCs measured by trypan blue exclusion assay were evaluated after 24 h of incubation.Results GCs cultured in group A and group B showed multiform morphology compared with other groups. GCs cultured in group D and group E were present with cytoplasmic atrophy and less pseudopodium. Moreover, group A and group B showed a similar level in the viability of GCs （P〉0.05）, which displayed no difference between group D and group E as well （P〉0.05）. Group C had a lower level of viability than group A （P〈0.05） but a higher level than group D （P〈0.01）. Conclusion Heat-inactivated sera can improve the growth of GCs. Different types of sera would have different effects on the growth of GCs cultured in vitro. The pre-culture with different types of sera should be performed to get better efficacy.
文摘Background： In the current society, infertility related to age has become a social problem. The in vitro fertilization （IVF） success rate in women with poor ovarian response （POR） is very low. Dandelion extract T-1 （DE-T1） is an effective component of the extract from the leaves and stems of Traxacum officinale, which is one of the medicines used in some patients with POR, but its molecular mechanism remains unclear. Methods： Following IVF, ovarian granulosa cells （GCs） of sixty patients were extracted and divided into normal ovarian response （NOR） and POR groups. GCs were cultured in a dose-dependent and time-dependent manner with DE-TI, proliferation of GCs was determined by Cell Counting Kit-8 assay, and mRNA levels of insulin-like growth factor 1 receptor （IGF-1R）, luteotropic hormone receptor （LHR）, follicle-stimulating hormone receptor （FSHR）, LHR, and CYP19A1 （aromatase） were determined by quantitative polymerase chain reaction. Progesterone and estradiol （E2） concentrations were determined by enzyme-linked immunosorbent assay. Results： The cell viability gradually increased with the progressive increase in the DE-T1 concentration. Compared with the control group （without DE-T1）, the mRNA expressions of FSHR, LHR, IGF-IR, and CYPIgAI were upregulated after the addition of DE-T l, especially in the 2.5% DE-T 1 group （P 〈 0.01 ）. The expression of IGF- 1R was upregulated approximately 25 times （24.97 ± 4.02 times） in the POR group with 2.5% DE-T1. E2 and progesterone levels increased with the increasing DE-T1 concentration. There were highly significant differences in the E2 and progesterone secretion between the NOR and POR groups （P 〈 0.01）. Conclusion： DE-TI may promote steroid hormone synthesis by promoting GC proliferation and upregulating GC receptor expression, thereby improving ovarian endocrine function.
