태아 고환에서 버팀세포의 미세형태학적 연구

이태진,윤삼현,김미경,박언섭,유재형,Lee, Tae-Jin,Yoon, Sam-Hyun,Kim, Mi-Kyung,Park, Eon-Sub,Yoo, Jae-Hyung 한국현미경학회 2001 Applied microscopy Vol.31 No.2

정상 성인 고환의 버팀세포(Sertoli cell)는 비분열세포이며, 정세관(seminiferous tubule)단면에서 비교적 불분명하게 관찰되고, 정세관 세포 성분의 $10\sim15%$를 차지하고 있다. 전자현미경적으로 버팅 세포는 특징적인 핵소체와 원형질막 및 세포질 소기관을 갖고 있다. 원형질막은 사춘기에 발달한 두 종류 즉 버팅세포와 버팀세포 및 버팅세포와 생식세포 사이의 세포연접을 가지고 있다. 그러나 태이에서 버팅세포의 정확한 미세구조에 대한 기술은 드물다. 이에 본 저자는 태아 고환의 발생 제 14주부터 제27주 사이의 17예를 수집하여 정상 미세구조를 확인하고, 태아기 버팀세포의 분화 양상을 알아보고자 하였다. 태아기에서 버팀세포와 생식세포 및 버팀세포와 버팀세포 사이의 세포연접은 부착반점과 비슷한 구조로 이루어져 있었고, 이들은 관찰 대상인 태령 제14주부터 관찰되었다. 태아기 버팅세포의 세포소기관의 발달은 전반적으로 미약하였다. 비교적 풍부하게 사립체가 태령 제14주부터 관찰되었고, 무과립세포질세망이 소수, 그리고 과립세포질세망이 비교적 풍부하게 관찰되었다. 지방소포의 수는 비교적 일정하게 관찰되었고, 포도당입자는 발생 단계에 따라 점차 증가하는 소견을 보였다. 미세섬유와 Charcot-Bottcher의 결정소체는 본 연구대상에서는 관찰되지 않았다. 결론적으로, 태아기의 버팀세포에서는 어른에서 관찰되는 특징적인 소견들이 관찰되지 않았으며, 어른과는 다소 다른 전자현미경 소견을 나타냈다. 하지만 버팅세포의 분화양상을 정확히 알기 위해서는 태령 제27주 이후부터 사춘기까지의 연구가 추가되어야 할 것으로 생각한다. Sertoli cells in the normal adult testis are nondividing cells, which are relatively inconspicuous on cross section of the seminiferous tubule and comprise about 10% to 15% of the tubular cellular elements. Ultrastructurally, Sertoli cells have characteristic nucleoli, plasma membrane, and cytoplasmic components. The plasma membrane has two types of intercellular junctions which are developed at puberty: junctions between adjacent Sertoli cells and Sertoli cell-germ ceil junction. However, the ultrastructural findings of Sertoli cells in human fetus is not fully elucidate yet. In the present study, human fetal testes ($14\sim27$ weeks) obtained from artificially induced abortions legally without gross malformation were studied using transmission electron microscopy to make clear the differentiation process of Sertoli cells in human. In human fetal testes from 14 weeks to 27 weeks, the cell junctions of Sertoli-germ cells and Sertoli-Sertoli cells are desmosome like structure and not tight junction or desmosome. The Overall intracytoplasmic organelles of Sertoli cells are relatively sparse. The mitochondrias are relatively abundant but no developed cristae. And the rough endoplasmic reticuli are abundant and smooth endoplasmic reticuli are sparse. The amount of lipid droplets are regularly observed in human fetal Sertoli cells. No microfilaments or Charcot-Bottcher's crystalloids are present. From the results, Sertoli cells in human fetal testes are somewhat different ultrastructural findings with puberty or adult. However, to make clear the differentiation process of Sertoli cells in human, further study for 28 weeks to puberty is required.

진도견(珍島犬)의 정자형성(精子形成)과 Sertoli세포(細胞) 특수(特殊) 연접부(連接部)의 미세구조(微細構造) II, Sertoli 세포(細胞) 특수(特殊) 연접부(連接部)의 미세구조(微細構造)

박영석,이재홍,Park, Young-seok,Lee, Jae-hong 대한수의학회 1992 大韓獸醫學會誌 Vol.32 No.3

In order to study on the Sertoli cell, we attempt have been made to measure the average number of each germ cells per Sertoli cell on the 12 stages of cycle in matured korean Jindo dog. The fine structure of Sertoli cell junctional specialization was studied with electron microscope. The results were summarized as follows; 1. The average number of various germ cells associated with Sertoli cell was 9.77 to 13. 80 through stages of cycle and the total average number was 11.62. 2. Sertoli-Sertoli cell junctional specialization was present in seminiferous epilthelium, and Sertoli-spermatid cell junctional specialization rose from stage 8 spermatid, persisted to step 13 spermatid and then disappeared. The structure of Sedoli-spermatid cell juncticnal specialization was not similar to that of Sertoli cxlls. 3. Just after spermiation, free-surface of Sertoli-spermatid cell junctional specialization was replaced by Sertoli cell cytoplasm with tubulobulbar complex at the neiglaboring region observed. 4. The Sertoli cell process was located within the cytoplasm of late stage spermatids. Some membranes of residual body and spermatid cytoplasm partly disappeared, resulting in opening of the cytoplasm of spermatid into that Sertoli cell. This fact suggested that spermatid cytoplasm was partly eliminated.

Changes in cAMP Response and Testosterone to 17β-Estradiol to FSH in Isolated Rat Sertoli Cells-Age and Dose Related Differences

Cho, Sung Hoon,Lee, Byung Churl,Seong, Sang Min CATHOLIC MEDICAL CENTER 1986 Bulletin of the Clinical Research Institute Vol.14 No.1

The Seroli cell was identified as> the main primary target site for the action of follicle stimulating hormone (FSH) in the testis. FSH has been known to bind specifically to Sertoli cell and to evoke a series of biochemical responses such as increased accumulation of cyclic AMP, production of androgen binding protein, and steroid metabolism. Recently age related changes in FSH stimulation of cAMP and aromatization of testosterone of 17β-estradiol were demonstrated. The purpose of this study was to investigate the changes in cAMP and testosterone to 17β-estradiol aromatization responses of FSH with dose differences in isolated rat Sertoli cells of different age. Methods for preparation of rat Sertoli cells were adapted from those used to prepare rat Sertoli cells. Homogenous preparations of primary Sertoli cell cultures were obtained from the testis of the rat of different age: prepubertal (14 days old rat), pubertal (18 days old rat) and adult (60 days old rat). For cAMP determinations, cells were stimulated for 30 min at 37?C with different dose of FSH in media containing 0.1 mg/m]l l-methyl-3-isobutylxanthine. cAMP assays were performed in duplicate using radioimmunoassay kit. FSH stimulated aromatization of testosterone of 17j3-estradiol was determined in cells stimulated with different dose of FSH in media containing 0.5 μmol/ml testosterone. 170-estradiol assays were performed in duplicate by means of specific radioimmunoassay. The results were as follows: 1. Sertoli cells from adult rats responded to FSH stimulation with over 30-fold rise at 5 min. At 30 min, cAMP is significantly lower and down to baseline level by 60 min. 2. There were age-related differences in Sertoli cell cAMP response after 30 min of FSH stimulation. Sertoli cells from pubertal rats responded most sensitively to FSH with a dose response relationship. 3. The heighest aromatase enzyme activity in converting testosterone to 17(3-estradiol was found in pubertal rat Sertoli cells. Sertoli cells from pubertal rats demonstrated a dose-response of FSH stimulation of testosterone to estradiol conversion. In conclusion, these results demonstrate that the age and dose-related differences in rat Sertoli cell response to FSH, and also provide direct evidence that the Sertoli cells represent a primary target site for FSH activity in the testes.

Evaluation of Bisphenol a Induced Apoptosis in Sertoli Cell-lines

김지향,김진규,도병록,이창주,윤용달 한국발생생물학회 2005 발생과 생식 Vol.9 No.2

본 연구는 Leydig 세포주와 Sertoli 세포주상에 bisphenol A(BPA)와 diethylstilbestrol(DES)의 영향을 알아보고자 수행하였다. 세포 종류에 따른 BPA의 영향을 알아보기 위해, BPA의 농도별로 두 세포주에 처리하여 세포생존율을 비교하였다. Sertoli 세포주가 Leydig 세포주에 비해서 저농도의 BPA에서 생존율이 유의하게 감소되는 것을 확인할 수 있어, Sertoli 세포가 Leydig 세포주에 비해 BPA에 The present report aimed at evaluating the effect of bisphenol A(BPA) and diethylstilbestrol(DES) on Leydig or Sertoli cell-lines. To identify the differences in the susceptibility to BPA upon different cell-types, assay of the cell viability was done on TM3(Leydig cells) and TM4(Sertoli cells) cell-lines. The result indicates that Sertoli cells are more sensitive to low dose of BPA than Leydig cells. Also, the BPA- or DES-treated Sertoli cells showed a reduction of phospholipase D(PLD) activity identically. According to the confirmation of the mRNA expression of fas receptor and fas ligand in the BPA-treated cells, fas/fasL system activated by BPA will deliver the apoptosis signal onto Sertoli TM4 cells. However, Fas/FasL system was not activated in the DES-treated cells unlike the BPA-treated cells.

Optimal Milieu for Culturing Porcine Sertoli Cell

Jabed Md. Anower,Kamal Tania,Kim, Byung-Ki The Korean Society of Animal Reproduction 2006 Reproductive & developmental biology Vol.30 No.3

The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum(FBS), follicular stimulating hormone(FSH), epidermal growth factor(EGF) and insulin-transferrin-sodium selenite(ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco's modified Eagle's medium+Ham's F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco's modified Eagle's medium+Ham's F-12 medium supplemented with 1% FBS, FSH EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

Sertoli-Leydig Cell Tumor 1예

박성철,한도규,김미진,고민환,이태형 대한부인종양 콜포스코피학회 2003 Journal of Gynecologic Oncology Vol.14 No.1

Sertoli-Leydig cell tumor는 탈여성화와 남성화 같은 증상을 특징적으로 나타내는 드문 난소종양의 하나이다. Sertoli-Leydig cell tumor는 조직적학적으로 섬유육종에서 보이는 방추형 세포로 대부분 구성되어 있어 감별진단이 요구된다. 구성대개 일측성이며 악성화가 낮아 가임기 여성의 경우 환측 난소난관절제술 및 난소의 추적 검사가 치료 원칙이나 폐경기 여성에서는 전 자궁절제술 및 양측 난소 난관 절세술을 시행한다. 잔류 종양이 있는 경우나 재발된 경우에는 암화학요법을 시행하기도 한다. 예후는 비교적 양호하며 5년 생존율은 70-90%로 5년 이후에는 재발이 드문 것으로 알려져 있다. 본 저자는 고령의 남성화 증상이 없는 미분화형 거대 Sertoli-Leydig cell tumor를 경험하였기에 이를 보고하는 바이다. The Sertoli-Leydig cell tumor is a rare gonadal tumor of sex-cord type, similar to that seen in the various phase of testicular development in the male. This tumor is most commom type of all virilizing ovarian tumors and account for less than 0.5% of all primary ovarian tumors. The Sertoli-Leydig cell tumor was histiologically mimics fibrosarcoma because the tumor shows fascicles of spindle shaped cells resembling fibrosarcoma in most of the part. The only thing differentiates Sertoli-Leydig cell tumor from fibrosarcoma is tubule of Sertoli cells. Recently we experienced a case of poorly differentiated Sertoli-Leydig cell tumor without virilization and so we present it with brief review of literature.

Optimal Milieu for Culturing Porcine Sertoli Cell

Md, Anower Jabed,Tania Kamel,Byung-Ki Kim 한국동물생명공학회(구 한국동물번식학회) 2006 Reproductive & developmental biology Vol.30 No.3

The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum (FBS), follicular stimulating hormone (FSH), epidermal growth factor (EGF) and insulin-transferrin-sodium selenite (ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco’s modified Eagle’s medium + Ham’s F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco’s modified Eagle’s medium + Ham’s F-12 medium supplemented with 1% FBS, FSH, EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

Extracellular domain of V-set and immunoglobulin domain containing 1 (VSIG1) interacts with Sertoli cell membrane protein, while its PDZ-binding motif forms a complex with ZO-1

Ekyune Kim,Youngjeon Lee,Kyu-Tae Chang 한국발생생물학회 2010 한국발생생물학회 학술발표대회 Vol.29 No.-

Mammalian spermatogenesis takes place in the seminiferousepithelium, which is composed of Sertoli cells and germ cells. The interaction between spermatogenic and Sertoli cells as well as elongated spermatids and Sertoli cells is tightly regulated by junctional adhesion molecules (JAMs). JAMs, which are cell adhesion molecules, are known to play roles in various biological processes such as fertilization, neurogenesis, cancer progression, and spermatogenesis. Members of the JAM family have a unique structure: they contain an N-terminal signal peptide domain, immunoglobulin (Ig)-like domains, transmembrane and cytoplasmic tail domains, each of which has distinct functions. The extracellular Ig-like domains interact in a homophilic or heterophilic manner, whereas cytoplasmic tail domain mediates the tight junction assembly. Although members of the JAM family are exclusively present in or restricted to the testis, their precise roles in spermatogenesis and fertilization have not yet been completely explored. The functional roles of Nectin-2, Nectin-3, JAM-C, cell adhesion molecule1 (CADM1), coxsackie and adenovirus receptor (CAR) have been evaluated by analysis of null mutant mice. Unfortunately, CAR-deficient mice had an embryonic lethal phenotype; this demonstrates the importance of CAR in development, but its physiological role in spermatogenesis is not known. The loss of CADM1, Nectin-3 and JAM-C resulted in male infertility caused by loss of adhesion between germ and Sertoli cells. A variety of JAMs participate in the interaction between germ and Sertoli cells. Recently, human VSIG1 has been characterized, which was originally known as A34, as a new member of the JAM family; VSIG1 is composed of two extracellular Ig-like domains, a transmembrane domain, and a cytoplasmic domain. However, this molecule has not been functionally characterized, so this was one of the aims of our present study. RT-PCR and immunoblot analyses were used to study VSIG1 expression, VSIG1 was specifically expressed in testicular germ cells but not in sperm. Pull-down assay with glutathione S-transferase (GST) or His-fused first Ig and second Ig domains of VSIG1 and SDS-PAGE under mild non-reducing conditions demonstrated that VSIG1 functions as an in vitro homophilic adhesion molecule. Furthermore, cells expressing a deletion of the C-terminus of VSIG1 failed to interact with ZO-1, the central structural protein of the tight junction. These findings suggest mouse VSIG1 interacts with an unknown molecule in Sertoli cells via its extracellular domain, while its cytoplasmic domain is needed for binding to ZO-1. Thus, we suggest mouse VSIG1 may play an important role in spermatogenesis rather than fertilization by forming heterophilic complex with a molecule similar to JAM family.

남성생식세포 Sertoli cell에 미치는 복분자(覆盆子)의 항산화 효과

김영주,장문석,박성규,Kim, Young Joo,Chang, Mun Seog,Park, Seong Kyu 대한한의학방제학회 2018 大韓韓醫學方劑學會誌 Vol.26 No.2

Objectives : The purpose of this study was to examine the antioxidant effects of the extract of Rubi Fructus on TM4 Sertoli cells. Methods : The extract was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity and cell viability assays on Sertoli cells. In addition, hydrogen peroxide-induced oxidative stress on Sertoli cells were examined by MTT assay. The antioxidant enzyme of Cu/Zn SOD, Mn SOD, catalase protein expression on Sertoli cells were also measured. Results : The results showed that the extract scavenged DPPH radical dose-dependent manner. The extract showed no cytotoxicity at concentration of 1, 5, 10, 50, $100{\mu}g/ml$. The hydrogen peroxide-induced cytotoxicity of Sertoli cells was protected to 88.3% by the extract at concentration of $100{\mu}g/ml$. Cu/Zn SOD and Mn SOD protein expression were significantly increased on Sertoli cells, but catalase protein expression was not significantly changed. Conclusions : In conclusion, the extract of Rubi Fructus has antioxidant effects on Sertoli cells and protect male reproductive system against oxidative stress.

Benzo[a]pyrene Cytotoxicity Tolerance in Testicular Sertoli Cells Involves Aryl-hydrocarbon Receptor and Cytochrome P450 1A1 Expression Deficiencies

Kim, Jin-Tac,Park, Ji-Eun,Lee, Seung-Jin,Yu, Wook-Joon,Lee, Hye-Jeong,Kim, Jong-Min The Korean Society of Developmental Biology 2021 발생과 생식 Vol.25 No.1

Benzo[a]pyrene (B[a]P) is a potent carcinogen and is classified as an endocrine-disrupting chemical. In mammalian testes, Sertoli cells support spermatogenesis. Therefore, if these cells are negatively affected by exposure to xenotoxic chemicals, spermatogenesis can be seriously disrupted. In this context, we evaluated whether mouse testicular TM4 Sertoli cells are susceptible to the induction of cytotoxicity-mediated cell death after exposure to B[a] P in vitro. In the present study, while B[a]P and B[a]P-7,8-diol were not able to induce cell death, exposure to BPDE resulted in cell death. BPDE-induced cell death is accompanied by the activation of caspase-3 and caspase-7. Depolarization of the mitochondrial membrane and cytochrome c release from mitochondria were observed in benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE)-treated cells. These results indicate that TM4 cells are susceptible to apoptosis in a caspase-dependent manner. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that aryl hydrocarbon receptor (AhR) expression was almost undetectable in TM4 cells and that its expression was not altered after B[a]P treatment. This indicates that TM4 cells are nearly AhR-deficient. In TM4 cells, the CYP1A1 protein and its activity were not present. From these results, it is clear that AhR may be a prerequisite for CYP1A1 expression in TM4 cells. Therefore, TM4 cells can be referred to as CYP1A1-deficient cells. Thus, TM4 Sertoli cells are believed to have a rigid and protective cellular machinery against genotoxic agents. In conclusion, it is suggested that tolerance to B[a]P cytotoxicity is associated with insufficient AhR and CYP1A1 expression in testicular Sertoli cells.