Isolation, Culture and Characterization of Human Sertoli Cells by Flow Cytometry: Development of Procedure

نویسندگان

  • Mohammad Reza Lakpour
  • Samaneh Aghajanpour
  • Morteza Koruji
  • Abdolhossein Shahverdi
  • Mohammad Ali Sadighi-Gilani
  • Marjan Sabbaghian
  • Reza Aflatoonian
  • Majid Rajabian-Naghandar
چکیده

BACKGROUND The sertoli cells in the testis create unique and safe environment to protect seminiferous tubules from auto antigens and invading pathogens. These cells produce the survival factor of the blood-testis barrier and produce special materials such as androgen binding proteins and contribute to the coordinated action of spermatogenesis. Given that the sertoli cells play an essential role in spermatogenesis and the lack of these cells leads to the disruption of spermatogenesis, it is necessary to investigate the behavior and performance of these cells. To achieve this goal, the cells must first be extracted. The aim of this study was to develop a procedure to isolate, culture, and characterize human sertoli cells. METHODS In order to isolate the sertoli cells of azoospermia patients who underwent (testicular sperm extraction) TESE surgery, washing up and multi_stage enzyme digestion of single cells, culture on petri dishes impregnated with datura stramonium lectin agglutinin (DSA) were done and then the cells were passaged for several times and isolated. For more purification, flow cytometry method with FSH receptor antibody was used. Immunocytochemistry assays and Elisa test for identification of these cells were employed. RESULTS The purification method resulted in more than 97% purity. The nature of sertoli cells was confirmed by morphology evaluation, detecting anti-mullerian hormone in sertoli cell culture media and the presence of FSH receptor on sertoli cells. CONCLUSION This study introduced and applied a method to isolate, culture, and purify human sertoli cells with high purity which made possible further researches on these cells.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Isolation and Characterization of Human Induced Pluripotent Stem Cells-Derived Mesenchymal Progenitors

Purpose: Isolating human induced pluripotent stem cells (hiPS)-derived mesenchymal progenitors as a new source of mesenchymal cells which can differentiate into different lineages like adipose and bone. Materials and Methods: After 7 days of hiPS1 culture on matrigle coated dishes, spindle like cells around colonies were removed by cell scraper. These cells that had mesenchymal like morphology ...

متن کامل

P-33: Expression of Toll-Like Receptor 2-3 Genes in Human Sertoli Cells

Background: Toll-like receptors (TLRs) constitute a major part of innate immunity, which can distinguish pathogen associate molecular pattern. Sertoli cells create a special immunological niche that protects somniferous tubules from auto antigens and pathogens. These cells are the only somatic cells in somniferous that protect testis cells against pathogens. The purpose of this study was to eva...

متن کامل

P-34: Expression of Toll-like Receptor2-3 Genes in Sertoli Cells of Patients with Azoospermia

Background: Toll-like receptors (TLRs) constitute a major part of innate immunity, which can distinguish pathogen associate molecular pattern. Sertoli cells create a special immunological niche that protects somniferous tubules from auto antigens and pathogens. These cells are the only somatic cells in somniferous that protect testis cells against pathogens. The purpose of this study was to eva...

متن کامل

جداسازی و ایمنوفنوتایپینگ سلول‌های اپی‌تلیالی پرده آمنیون

Abstract Background: Human amniotic epithelial cells (hAECs) have been introduced as a valuable source of stem cells with therapeutic applications. Despite the extensive study and emerging evidence, cell surface marker signature of hAECs remained controversial. The aim of the present study was to establish an efficient and optimized isolation procedure of hAECs and their characterization in ter...

متن کامل

A Novel mAb against a Human CD34 Peptide Reacts with the Native Protein on CD34+ Cells

Background: Human CD34 is a transmembrane glycoprotein which is expressed in human hematopoietic stem cells (HSCs) and the small- vessel endothelial cells of a variety of tissues. CD34 plays a critical role as a marker for diagnosis and classification of leukemia. Anti CD34 antibodies are used for isolation and purification of HSCs from bone marrow, peripheral blood and cord blood. Objective: ...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 18  شماره 

صفحات  -

تاریخ انتشار 2017