Comparison of colony formation of human spermatogonial stem cells (SSCs) with and without collagen.
J Pak Med Assoc
; 66(3): 285-91, 2016 Mar.
Article
em En
| MEDLINE
| ID: mdl-26968278
ABSTRACT
OBJECTIVE:
To investigate the effects of collagen and growth factors on in vitro proliferation of human spermatogonial stem cells obtained from patients with non-obstructive azoospermia.METHODS:
The experimental cross-sectional study was conducted from February 2013 to April 2015 after obtaining approval from the ethics committee of Ahvaz Jundishapur University of Medical Sciences, Iran. Testicular sperm extractions of non-obstructive azoospermic patients were obtained from the Clinical Urology and Embryology, In Vitro Fertilization Department of Imam Khomeini Hospital. Spermatogonial stem cells and Sertoli cells, obtained from human testis biopsies by a two-step enzymatic digestion method, were purified using fluorescence- activated cell-sorting and daturastramonium-lectin, and were cultured separately. To investigate a more direct influential factor on colony formation, one control and two experimental groups were formed. Group 1 acted as the control in which spermatogonial stem cells were co-cultured with Sertoli cells alone. In group 2 they were co-cultured with Sertoli cells and growth factors such as leukaemia inhibitory factor, epidermal growth factor and glial cell-derived neurotrophic factor, and in group 3 with Sertoli cells along with growth factors in the presence of collagen-coated dishes. Number and diameter of the colonies were evaluated after 7 weeks.RESULTS:
Specimens obtained related to 21 patients. Number and diameter of the colonies in group 3 (18±2.6 and 276.6±45.5) were significantly more than both groups 1 (3.5±1 and D181.6±12) and group 2(11±2.2 and 165.2±32.5) (p<0.05 each). Also, the number and diameter of colony in group 2 were significantly better than the control group (p<0.05).Expression profile of the VASA, promyelocytic leukaemia zinc-finger (PLZF), Octamer-binding transcription factor 4 (OCT4) and integrin a6 (INTGa6) were detected in all groups. Based on cytochemical findings, OCT4 was expressed in the colonies of all three groups.CONCLUSIONS:
According to positive effects of collagen and growth factors on the colonisation of spermatogonial stem cells, it seems that using the cells may lead to better colonisation of this type of stem cells.Palavras-chave
Buscar no Google
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Colágeno
/
Proliferação de Células
/
Células-Tronco Germinativas Adultas
Tipo de estudo:
Observational_studies
/
Prevalence_studies
/
Risk_factors_studies
Aspecto:
Ethics
Limite:
Humans
/
Male
Idioma:
En
Revista:
J Pak Med Assoc
Ano de publicação:
2016
Tipo de documento:
Article