In vitro Proliferation of Chick Primordial Germ Cells Co-cultured with Germinal Ridge Stroma Cells
In order to develop the technique for in vitro culture of chicken primordial germ cells obtained from circulating embryonic blood vessels (cPGCs), participation of feeder cells and some of growth factors was examined. In this experiment, Dulbecco's MEM (DMEM) mixed with Ham's F12 w...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Japan Poultry Science Association
2001-04-01
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| Series: | The Journal of Poultry Science |
| Subjects: | |
| Online Access: | https://www.jstage.jst.go.jp/article/jpsa/38/2/38_2_152/_pdf/-char/en |
| _version_ | 1797817084084748288 |
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| author | Ryou Fukushima Noboru Fujihara |
| author_facet | Ryou Fukushima Noboru Fujihara |
| author_sort | Ryou Fukushima |
| collection | DOAJ |
| description | In order to develop the technique for in vitro culture of chicken primordial germ cells obtained from circulating embryonic blood vessels (cPGCs), participation of feeder cells and some of growth factors was examined. In this experiment, Dulbecco's MEM (DMEM) mixed with Ham's F12 was used in combination with some kind of cell growth factors such as chicken stem cells factor (SCF), human leukemia inhibitory factor (LIF), human basic fibroblast growth factor (bFGF) and chicken insulin-like growth factor (IGF-1). In the present study, chicken embryos incubated for 5, 6 and 7 days were subjected to the isolation of stroma cells from germinal ridges. These stroma cells isolated from germinal ridge which were used as feeder cells, were divided into two parts, left side stroma cells (L-GRSCs) and right ones (R-GRSCs). The cPGCs were obtained by the filtration method from blood samples aspirated from extra-embryonic veins. The short-term (6 days) culture of cPGCs with L-GRSCs derived from 5 days embryos showed slightly increased proliferation of the cells, by around 1.7-2.3 times. Contrariwise, the co-culture of cPGCs with R-GRSCs did not bring about any increase of PGCs number compared with the case of L-GRSCs. On the one hand, the co-culture with both L- and R-GRSCs derived from 6 and 7 day embryos, resulting in the disappearance of the cells after 7 days of culture. In case of the long-term (30 days) culture with L- and R-GRSCs, the number of PGCs significantly (P<0.05) increased, showing 70 to 100 times increment. In these cases, no significant difference was observed between L- and R-GRSCs or among growth factors added. In addition to this, the PGCs cultured for 30 days formed a kind of cell clumps surrounded by the co-cultured GRSCs. The results obtained in this experiment suggest the possibility of proliferation of the PGCs outside the body by co-culturing with germinal ridge stroma cells derived from the 5 day embryos. |
| first_indexed | 2024-03-13T08:47:27Z |
| format | Article |
| id | doaj.art-ce8a1b8c3a0144559f677c2769a9f150 |
| institution | Directory Open Access Journal |
| issn | 1346-7395 1349-0486 |
| language | English |
| last_indexed | 2024-03-13T08:47:27Z |
| publishDate | 2001-04-01 |
| publisher | Japan Poultry Science Association |
| record_format | Article |
| series | The Journal of Poultry Science |
| spelling | doaj.art-ce8a1b8c3a0144559f677c2769a9f1502023-05-30T00:07:00ZengJapan Poultry Science AssociationThe Journal of Poultry Science1346-73951349-04862001-04-0138215215910.2141/jpsa.38.152jpsaIn vitro Proliferation of Chick Primordial Germ Cells Co-cultured with Germinal Ridge Stroma CellsRyou Fukushima0Noboru Fujihara1Laboratory of Animal Reproductive Physiology, Division of Animal Resource Science, School of Agriculture, Graduate School Kyushu UniversityLaboratory of Animal Reproductive Physiology, Division of Animal Resource Science, School of Agriculture, Graduate School Kyushu UniversityIn order to develop the technique for in vitro culture of chicken primordial germ cells obtained from circulating embryonic blood vessels (cPGCs), participation of feeder cells and some of growth factors was examined. In this experiment, Dulbecco's MEM (DMEM) mixed with Ham's F12 was used in combination with some kind of cell growth factors such as chicken stem cells factor (SCF), human leukemia inhibitory factor (LIF), human basic fibroblast growth factor (bFGF) and chicken insulin-like growth factor (IGF-1). In the present study, chicken embryos incubated for 5, 6 and 7 days were subjected to the isolation of stroma cells from germinal ridges. These stroma cells isolated from germinal ridge which were used as feeder cells, were divided into two parts, left side stroma cells (L-GRSCs) and right ones (R-GRSCs). The cPGCs were obtained by the filtration method from blood samples aspirated from extra-embryonic veins. The short-term (6 days) culture of cPGCs with L-GRSCs derived from 5 days embryos showed slightly increased proliferation of the cells, by around 1.7-2.3 times. Contrariwise, the co-culture of cPGCs with R-GRSCs did not bring about any increase of PGCs number compared with the case of L-GRSCs. On the one hand, the co-culture with both L- and R-GRSCs derived from 6 and 7 day embryos, resulting in the disappearance of the cells after 7 days of culture. In case of the long-term (30 days) culture with L- and R-GRSCs, the number of PGCs significantly (P<0.05) increased, showing 70 to 100 times increment. In these cases, no significant difference was observed between L- and R-GRSCs or among growth factors added. In addition to this, the PGCs cultured for 30 days formed a kind of cell clumps surrounded by the co-cultured GRSCs. The results obtained in this experiment suggest the possibility of proliferation of the PGCs outside the body by co-culturing with germinal ridge stroma cells derived from the 5 day embryos.https://www.jstage.jst.go.jp/article/jpsa/38/2/38_2_152/_pdf/-char/enchickenpgcsin vitro cultureproliferationgerminal ridge stroma cells |
| spellingShingle | Ryou Fukushima Noboru Fujihara In vitro Proliferation of Chick Primordial Germ Cells Co-cultured with Germinal Ridge Stroma Cells The Journal of Poultry Science chicken pgcs in vitro culture proliferation germinal ridge stroma cells |
| title | In vitro Proliferation of Chick Primordial Germ Cells Co-cultured with Germinal Ridge Stroma Cells |
| title_full | In vitro Proliferation of Chick Primordial Germ Cells Co-cultured with Germinal Ridge Stroma Cells |
| title_fullStr | In vitro Proliferation of Chick Primordial Germ Cells Co-cultured with Germinal Ridge Stroma Cells |
| title_full_unstemmed | In vitro Proliferation of Chick Primordial Germ Cells Co-cultured with Germinal Ridge Stroma Cells |
| title_short | In vitro Proliferation of Chick Primordial Germ Cells Co-cultured with Germinal Ridge Stroma Cells |
| title_sort | in vitro proliferation of chick primordial germ cells co cultured with germinal ridge stroma cells |
| topic | chicken pgcs in vitro culture proliferation germinal ridge stroma cells |
| url | https://www.jstage.jst.go.jp/article/jpsa/38/2/38_2_152/_pdf/-char/en |
| work_keys_str_mv | AT ryoufukushima invitroproliferationofchickprimordialgermcellscoculturedwithgerminalridgestromacells AT noborufujihara invitroproliferationofchickprimordialgermcellscoculturedwithgerminalridgestromacells |