Differentiation of Human Bone Marrow Mesenchymal Stem Cells into Leydig or Steroidogenic Cells in vivo

REN Shang-qing, DENG Shi, WU Yun-jian. et al

Abstract

To study the differentiation of human bone marrow mesenchymal stem cells (BMSCs) into Leydig or steroidogenic cells in vivo and the immunoreaction related to transplantation into mouse testis. Methods After differentiation and cultivation, the 3 rd-passage BMSCs were collected and labeled with Hoechest 33342, and joined the saline fluorouracil to form cell-suspending fluid. After injection of the etgane dimethane sulphonate (EDS), the mice received the transplanted cell-suspending fluid by testis net injection with a dose of each side testicular 0.05 mL. Since the first day prior to transplantation, mice were executed every 2 d (one mouse each time) and the testosterone concentrations were analyzed. The control group included 20 BALB/c mice without any treatment during the same period. The results were analyzed by microscopic observation, using 3β-hydroxysteroid dehydrogenase (3β-HSD) monoclonal antibody and mouse anti-human cell nucleus monoclonal antibody for immunofluorescence assay on the testis. For tracking the BMSCs, the cells which were positively stained with both 3β-HSD and mouse anti-human cell nucleus monoclonal antibodies were retrieved. Results Certain killing effect of EDS to the mouse Leydig cells was observed. Transplantation of human BMSCs into the mouse testis by testis net injection was effective and feasible, no immunoreactions were detected. After transplantation, no positive cells of 3β-HSD and mouse anti-human cell nucleus monoclonal antibody were found. Conclusion Transplantation of human BMSCs into the mouse testis by testis net injection was effective and feasible, no immunoreactions were detected. After transplantation, the human BMSCs failed to differentiate into Leydig cells or steroidogenic cells.

 

Keywords: Mesenchymal stem cells, Leydig cells, Testicle 

 

Full Text:

PDF


References


Wang XY, Xing X. Zhou GD, et al. Testosterone-secreting tissue construction with tissue engineering technique. Plast Reconstr Surg.2007; 120( 1):34-36.

Chen H, Ge RS, Zirkin BR. Leydig cells; from stem cells to aging. Mol Cell Endocrinol,2009,306(1-2) ;9-16.

Erices A, Conget P. Minguell JJ. Mesenchymal progenitor cells in human umbilical cord blood. Br J Haematol, 2000; 109 ( 1 ); 235-224. 201

De Bari С, Dell’ Accio F, Tylzanowski Р, et al. Multipotent mesenchymal stem cells from adult human synovial membrane. Arthritis Rheum,2001; 44(8): 1928-1942.

Maitra B, Szekely E, Gjini K, et al. Human mesenchymal stem cells support unrelated donor hematopoietic stem cells and suppress T-cell activation. Bone Marrow Transplant. 2004» 33 (6): 597-604.

Yazawa T, Mizutani T, Yamada K. et al. Differentiation of adult stem cells derived from hone marrow stroma into Leydig or adrenocortical cells. Endocrinology, 2006; 147 (9); 4104- 4111.

Gondo S, Okabe T, Tanaka T, et al. Adipose tissue-derived and bone marrow-derived mesenchymal cells develop into different lineage of steroidogenic cells by forced expression of steroidogenic factor 1. Endocrinology, 2008; 149 (9); 4717- 4725.

Sriraman V, Sairam MR. Rao AJ. Evaluation of relative roles of LH and FSH in regulation of differentiation of Leydig cells using an ethane 1, 2-dimethylsulfonate-treated adult rat model. J Endocrinol,2003; 176(1): 151-1611.

Morris ID. Leydig cell resistance to the cytotoxic effect of ethylene dimethanesulphonate in the adult rat testis. J Endocrinol, 1985; 105(3); 311-316.

Bakalska M, Atanassova N, Koeva Y, et al. Induction of male germ cell apoptosis by testosterone withdrawal after ethane dimethanesulfonate treatment in adult rats. Endocr Regul. 2004;38(3):103-110.

Jackson AE, O’Leary PC, Ayers MM, et al. The effects of ethylene dimethane sulphonate (EDS) on rat Leydig cells; evidence to support a connective tissue origin of Leydig cells. Biol Reprod, 1986;35(2) ;425-437.

Gabory A, Jammes H. Dandolo L. The H19 locus; role of an imprinted non-coding RNA in growth and development. Bioessays, 2010; 32(6); 4 73-480.

Nativio R, Wendt KS, Ito Y, et al. Cohesin is required for higher-order chromatin conformation at the imprinted IGF2- H19 locus. PLoS Genet,2009;5( 11) :el000739.

Yao H, Brick K, Evrard Y, et al. Mediation of CTCF transcriptional insulation by DEAD-box RNA-binding protein p68 and steroid receptor RNA activator SRA. Gene Dev,2010; 24(22):2543-2555.

Ratajczak MZ. Igf2-H19. an imprinted tandem gene’ is an important regulator of embryonic development, a guardian of proliferation of adult pluripotent stem cells, a regulator of longevity, and a *passkey* to cancerogenesis. Folia Histochem Cytobiol,2012; 50( 2); 171-179.

Wu J, Qin Y, Li B, et al. Hypomethylated and hypermethylated profiles of H19DMR are associated with the aberrant imprinting of IGF2 and H19 in human hepatocellular carcinoma. Genomics, 2008 ;91(5):443-450.

Wan LB. Pan H. Hannenhalli S. et al. Maternal depletion of CTCF reveals multiple functions during oocyte and preimplantation embryo development. Development, 2008; 135 (16):2729-2738.

Heath H, De Almeida CR, Sleutels F, elal. CTCF regulates cell cycle progression of a{3 T cells in the thymus. EMBO J, 2008;27(21);2839-2850.

Torrano V, Chernukhin I, Docquier F, et al. CTCF regulates growth and erythroid differentiation of human myeloid leukemia cells. J Biol Chem, 2005; 280(30); 28152-28161.

Rasko JEJ, Klenova P3M, Leon J, et al. Cell growth inhibition by the multifunctional multivalent zinc-finger factor CTCF. Cancer Res,2001 ;61( 16) ;6002-6007.


Refbacks

  • There are currently no refbacks.