Isolation and characterization of human spermatogonial stem cells
<p>Abstract</p> <p>Background</p> <p>To isolate and characterization of human spermatogonial stem cells from stem spermatogonium.</p> <p>Methods</p> <p>The disassociation of spermatogonial stem cells (SSCs) were performed using enzymatic digestio...
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| Format: | Article |
| Language: | English |
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BMC
2011-10-01
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| Series: | Reproductive Biology and Endocrinology |
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| Online Access: | http://www.rbej.com/content/9/1/141 |
| _version_ | 1818083977946202112 |
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| author | Liu Shixue Tang Ziwei Xiong Tao Tang Wei |
| author_facet | Liu Shixue Tang Ziwei Xiong Tao Tang Wei |
| author_sort | Liu Shixue |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>To isolate and characterization of human spermatogonial stem cells from stem spermatogonium.</p> <p>Methods</p> <p>The disassociation of spermatogonial stem cells (SSCs) were performed using enzymatic digestion of type I collagenase and trypsin. The SSCs were isolated by using Percoll density gradient centrifugation, followed by differential surface-attachment method. Octamer-4(OCT4)-positive SSC cells were further identified using immunofluorescence staining and flow cytometry technques. The purity of the human SSCs was also determined, and a co-culture system for SSCs and Sertoli cells was established.</p> <p>Results</p> <p>The cell viability was 91.07% for the suspension of human spermatogonial stem cells dissociated using a two-step enzymatic digestion process. The cells isolated from Percoll density gradient coupled with differential surface-attachement purification were OCT4 positive, indicating the cells were human spermatogonial stem cells. The purity of isolated human spermatogonial stem cells was 86.7% as assessed by flow cytometry. The isolated SSCs were shown to form stable human spermatogonial stem cell colonies on the feeder layer of the Sertoli cells.</p> <p>Conclusions</p> <p>The two-step enzyme digestion (by type I collagenase and trypsin) process is an economical, simple and reproducible technique for isolating human spermatogonial stem cells. With little contamination and less cell damage, this method facilitates isolated human spermatogonial stem cells to form a stable cell colony on the supporting cell layer.</p> |
| first_indexed | 2024-12-10T19:46:34Z |
| format | Article |
| id | doaj.art-4770484481464e0a83d5edc8f28466ad |
| institution | Directory Open Access Journal |
| issn | 1477-7827 |
| language | English |
| last_indexed | 2024-12-10T19:46:34Z |
| publishDate | 2011-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Reproductive Biology and Endocrinology |
| spelling | doaj.art-4770484481464e0a83d5edc8f28466ad2022-12-22T01:35:52ZengBMCReproductive Biology and Endocrinology1477-78272011-10-019114110.1186/1477-7827-9-141Isolation and characterization of human spermatogonial stem cellsLiu ShixueTang ZiweiXiong TaoTang Wei<p>Abstract</p> <p>Background</p> <p>To isolate and characterization of human spermatogonial stem cells from stem spermatogonium.</p> <p>Methods</p> <p>The disassociation of spermatogonial stem cells (SSCs) were performed using enzymatic digestion of type I collagenase and trypsin. The SSCs were isolated by using Percoll density gradient centrifugation, followed by differential surface-attachment method. Octamer-4(OCT4)-positive SSC cells were further identified using immunofluorescence staining and flow cytometry technques. The purity of the human SSCs was also determined, and a co-culture system for SSCs and Sertoli cells was established.</p> <p>Results</p> <p>The cell viability was 91.07% for the suspension of human spermatogonial stem cells dissociated using a two-step enzymatic digestion process. The cells isolated from Percoll density gradient coupled with differential surface-attachement purification were OCT4 positive, indicating the cells were human spermatogonial stem cells. The purity of isolated human spermatogonial stem cells was 86.7% as assessed by flow cytometry. The isolated SSCs were shown to form stable human spermatogonial stem cell colonies on the feeder layer of the Sertoli cells.</p> <p>Conclusions</p> <p>The two-step enzyme digestion (by type I collagenase and trypsin) process is an economical, simple and reproducible technique for isolating human spermatogonial stem cells. With little contamination and less cell damage, this method facilitates isolated human spermatogonial stem cells to form a stable cell colony on the supporting cell layer.</p>http://www.rbej.com/content/9/1/141spermatogonial stem cellsspermatogenesisspermatogoniaspermatocytessperm cellsmale infertilityandrology |
| spellingShingle | Liu Shixue Tang Ziwei Xiong Tao Tang Wei Isolation and characterization of human spermatogonial stem cells Reproductive Biology and Endocrinology spermatogonial stem cells spermatogenesis spermatogonia spermatocytes sperm cells male infertility andrology |
| title | Isolation and characterization of human spermatogonial stem cells |
| title_full | Isolation and characterization of human spermatogonial stem cells |
| title_fullStr | Isolation and characterization of human spermatogonial stem cells |
| title_full_unstemmed | Isolation and characterization of human spermatogonial stem cells |
| title_short | Isolation and characterization of human spermatogonial stem cells |
| title_sort | isolation and characterization of human spermatogonial stem cells |
| topic | spermatogonial stem cells spermatogenesis spermatogonia spermatocytes sperm cells male infertility andrology |
| url | http://www.rbej.com/content/9/1/141 |
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