Spermatogonial stem cell enrichment using simple grafting of testis and in vitro cultivation

نویسندگان

  • Jung Jin Lim
  • Dong Won Seol
  • Kyung Hee Choi
  • Dong Hyuk Shin
  • Hyung Joon Kim
  • Seung-Hun Song
  • Dong Ryul Lee
چکیده

Enrichment of spermatogonial stem cells (SSCs) from the mammalian adult testis faces several limitations owing to their relatively low numbers among many types of advanced germ cells and somatic cells. The aim of the present study was to improve the isolation efficiency of SSCs using a simple tissue grafting method to eliminate the existing advanced germ cells. Sliced testis parenchyma obtained from adult ICR or EGFP-expressing transgenic mice were grafted heterotropically under the dorsal skin of nude mice. The most advanced germ cells disappeared in the grafted tissues after 2-4 weeks. Grafted tissues were dissociated enzymatically and plated in culture dishes. During in vitro culture, significantly more SSCs were obtained from the grafted testes than from non-grafted controls, and the isolated SSCs had proliferative potential and were successfully maintained. Additionally, EGFP-expressing SSCs derived from graft parenchyma were transplanted into bulsufan-treated recipient mice testes. Finally, we obtained EGFP-expressing pups after in vitro fertilization using spermatozoa derived from transplanted SSCs. These results suggest that subcutaneous grafting of testis parenchyma and the subsequent culture methods provide a simple and efficient isolation method to enrich for SSCs in adult testis without specific cell sorting methods and may be useful tools for clinical applications.

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

Combination of In Vivo Cryptorchid Testis and In Vitro Co- Culture System to Obtain High Purification and Proliferation of Mouse Spermatogonial Stem Cells

Background The present study was designed to evaluate the survival and proliferation of spermatogonial stem cells from cryptorchid mouse testis in co-culture system over a 3 weeks period. MaterialsAndMethods Sertoli and spermatogonial cells were isolated from bilateral cryptorchid mouse model testes. Isolated spermatogonial cells were co-cultured with Sertoli cells in minimal essential medium (...

متن کامل

Establishment of Oxidative Stress Modeling during Spermatogonial Stem Cells Cultivation Treated with Different Doses of H2O2

Introduction: Nowadays, spermatogonial stem cells (SSCs) cultivation has been used by many researchers as an effective tool for infertility treatments. Oxidative conditions can be effective on cell proliferation and differentiation of these cells. So, the aim of this study was to establish oxidative stress model for antioxidant activity of some drugs investigation during SSCs in vitro culture. ...

متن کامل

Evaluation of the Effect of Follicle Stimulating Hormone (FSH) on Survival and Colonization of Caprine Spermatogonial Stem Cells during in Vitro Culture

Background and Objectives: Spermatogonial stem cells are specific cells that have the ability of self-renewal and differentiation. These cells play an essential role in maintaining spermatogenesis and fertility. In this regard, the present study was performed with the purpose of investigating the effect of different concentrations of follicle stimulating hormone (FSH) on in vitro colony formati...

متن کامل

O-3: Identification and Characterization of Repopulating Spermatogonial Stem Cells from The Adult Human Testis

Background: This study was conducted to identify and characterize repopulating spermatogonial stem cells (SSCs) in the adult human testes. Materials and Methods: Testes biopsies from obstructive azoospermic patients and normal segments of human testicular tissue were used. Flow cytometry, real time PCR and immunohistochemical analysis were performed. Purified human spermatogonia were transplant...

متن کامل

Spermatogonial stem cell enrichment by multiparameter selection of mouse testis cells.

The spermatogonial stem cell initiates and maintains spermatogenesis in the testis. To perform this role, the stem cell must self replicate as well as produce daughter cells that can expand and differentiate to form spermatozoa. Despite the central importance of the spermatogonial stem cell to male reproduction, little is known about its morphological or biochemical characteristics. This result...

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 4  شماره 

صفحات  -

تاریخ انتشار 2014