ALK Family Inhibitor A83-01 Promotes the Proliferation of Mouse Male Germline Stem Cells (mGSCs) Under Serum- and Feeder-Free Conditions
A83-01 is a selective inhibitor of the TGF-β type I receptor ALK, which inhibits the TGF-β-induced epithelial-to-mesenchymal transition (EMT) via the inhibition of Smad2 phosphorylation. Previous studies have showed that A83-01 promoted somatic cellular reprogramming significantly. Male germline ste...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
KeAi Communications Co., Ltd.
2013-10-01
|
| Series: | Journal of Integrative Agriculture |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S209531191360413X |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849251090297520128 |
|---|---|
| author | Meng YU Long WANG Yue HU Zhi-min LIAN Jin-lian HUA |
| author_facet | Meng YU Long WANG Yue HU Zhi-min LIAN Jin-lian HUA |
| author_sort | Meng YU |
| collection | DOAJ |
| description | A83-01 is a selective inhibitor of the TGF-β type I receptor ALK, which inhibits the TGF-β-induced epithelial-to-mesenchymal transition (EMT) via the inhibition of Smad2 phosphorylation. Previous studies have showed that A83-01 promoted somatic cellular reprogramming significantly. Male germline stem cells (mGSCs), as an alternative resource of pluripotent stem cells derived adult testis, have promising valuable in clinic medicine and regeneration, however, the derivation of mGSCs was complex and difficult. What the role A83-01 plays in promoting the proliferation of mGSCs is still unknown. In this study, combined with A83-01 and knockout serum replacement (KSR) medium, we obtained a relatively feeder- and serum-free system for mGSCs culturing in vitro and the optimal concentration of A83-01 was 0.25 μmol L−1. After continuous culturing, the proliferation efficiency of undifferentiated mGSCs and differentiation capacity of mGSC were examined as well. Results showed that, A83-01 dramatically increased the number of mGSCs and AP positive colonies, and the mitosis index according to the BrdU assay. A83-01 could also increase the expression of pluripotent markers including Oct4, Klf4, Nanog and c-Myc, analyzed by real-time quantative PCR. mGSCs cultured in the optimal feeder-and serum-free system combined with A83-01 could form embryoid bodies (EBs), which consisted of three embryonic layers detected by immunofluorescence and RT-PCR. Remarkably, the results demonstrated 0.25 μmol L−1 A83-01 could promote the proliferation of mouse mGSC colonies and maintain their undifferentiated status under feeder- and serum-free systems. |
| format | Article |
| id | doaj-art-9f9c215914a547f48ff557d40b1600b2 |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2013-10-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-9f9c215914a547f48ff557d40b1600b22025-08-20T03:57:03ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192013-10-0112101839184610.1016/S2095-3119(13)60413-XALK Family Inhibitor A83-01 Promotes the Proliferation of Mouse Male Germline Stem Cells (mGSCs) Under Serum- and Feeder-Free ConditionsMeng YU0Long WANG1Yue HU2Zhi-min LIAN3Jin-lian HUA4College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology/Key Lab for Animal Biotechnology, Agriculture Ministry of China/Northwest A&F University, Yangling 712100, P.R. ChinaCollege of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology/Key Lab for Animal Biotechnology, Agriculture Ministry of China/Northwest A&F University, Yangling 712100, P.R. ChinaCollege of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology/Key Lab for Animal Biotechnology, Agriculture Ministry of China/Northwest A&F University, Yangling 712100, P.R. ChinaCollege of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology/Key Lab for Animal Biotechnology, Agriculture Ministry of China/Northwest A&F University, Yangling 712100, P.R. ChinaCorrespondence HUA Jin-lian, Tel: +86-29-87080068, Fax: +86-29-87080068; College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology/Key Lab for Animal Biotechnology, Agriculture Ministry of China/Northwest A&F University, Yangling 712100, P.R. ChinaA83-01 is a selective inhibitor of the TGF-β type I receptor ALK, which inhibits the TGF-β-induced epithelial-to-mesenchymal transition (EMT) via the inhibition of Smad2 phosphorylation. Previous studies have showed that A83-01 promoted somatic cellular reprogramming significantly. Male germline stem cells (mGSCs), as an alternative resource of pluripotent stem cells derived adult testis, have promising valuable in clinic medicine and regeneration, however, the derivation of mGSCs was complex and difficult. What the role A83-01 plays in promoting the proliferation of mGSCs is still unknown. In this study, combined with A83-01 and knockout serum replacement (KSR) medium, we obtained a relatively feeder- and serum-free system for mGSCs culturing in vitro and the optimal concentration of A83-01 was 0.25 μmol L−1. After continuous culturing, the proliferation efficiency of undifferentiated mGSCs and differentiation capacity of mGSC were examined as well. Results showed that, A83-01 dramatically increased the number of mGSCs and AP positive colonies, and the mitosis index according to the BrdU assay. A83-01 could also increase the expression of pluripotent markers including Oct4, Klf4, Nanog and c-Myc, analyzed by real-time quantative PCR. mGSCs cultured in the optimal feeder-and serum-free system combined with A83-01 could form embryoid bodies (EBs), which consisted of three embryonic layers detected by immunofluorescence and RT-PCR. Remarkably, the results demonstrated 0.25 μmol L−1 A83-01 could promote the proliferation of mouse mGSC colonies and maintain their undifferentiated status under feeder- and serum-free systems.http://www.sciencedirect.com/science/article/pii/S209531191360413Xmale germline stem cells (mGSCs)A83-01feeder- and serum-freemouse |
| spellingShingle | Meng YU Long WANG Yue HU Zhi-min LIAN Jin-lian HUA ALK Family Inhibitor A83-01 Promotes the Proliferation of Mouse Male Germline Stem Cells (mGSCs) Under Serum- and Feeder-Free Conditions Journal of Integrative Agriculture male germline stem cells (mGSCs) A83-01 feeder- and serum-free mouse |
| title | ALK Family Inhibitor A83-01 Promotes the Proliferation of Mouse Male Germline Stem Cells (mGSCs) Under Serum- and Feeder-Free Conditions |
| title_full | ALK Family Inhibitor A83-01 Promotes the Proliferation of Mouse Male Germline Stem Cells (mGSCs) Under Serum- and Feeder-Free Conditions |
| title_fullStr | ALK Family Inhibitor A83-01 Promotes the Proliferation of Mouse Male Germline Stem Cells (mGSCs) Under Serum- and Feeder-Free Conditions |
| title_full_unstemmed | ALK Family Inhibitor A83-01 Promotes the Proliferation of Mouse Male Germline Stem Cells (mGSCs) Under Serum- and Feeder-Free Conditions |
| title_short | ALK Family Inhibitor A83-01 Promotes the Proliferation of Mouse Male Germline Stem Cells (mGSCs) Under Serum- and Feeder-Free Conditions |
| title_sort | alk family inhibitor a83 01 promotes the proliferation of mouse male germline stem cells mgscs under serum and feeder free conditions |
| topic | male germline stem cells (mGSCs) A83-01 feeder- and serum-free mouse |
| url | http://www.sciencedirect.com/science/article/pii/S209531191360413X |
| work_keys_str_mv | AT mengyu alkfamilyinhibitora8301promotestheproliferationofmousemalegermlinestemcellsmgscsunderserumandfeederfreeconditions AT longwang alkfamilyinhibitora8301promotestheproliferationofmousemalegermlinestemcellsmgscsunderserumandfeederfreeconditions AT yuehu alkfamilyinhibitora8301promotestheproliferationofmousemalegermlinestemcellsmgscsunderserumandfeederfreeconditions AT zhiminlian alkfamilyinhibitora8301promotestheproliferationofmousemalegermlinestemcellsmgscsunderserumandfeederfreeconditions AT jinlianhua alkfamilyinhibitora8301promotestheproliferationofmousemalegermlinestemcellsmgscsunderserumandfeederfreeconditions |