Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity
Fetal cells are widely considered a superior cell source for regenerative medicine; fetal cells show higher proliferative capacity and have undergone fewer replicative cycles that could generate spontaneous mutations. Fetal cells in amniotic fluid were among the first normal primary cells to be cult...
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| Format: | Article |
| Language: | English |
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Wiley
2012-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2012/485950 |
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| _version_ | 1832548165842108416 |
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| author | Patricia G. Wilson Lorna Devkota Tiffany Payne Laddie Crisp Allison Winter Zhan Wang |
| author_facet | Patricia G. Wilson Lorna Devkota Tiffany Payne Laddie Crisp Allison Winter Zhan Wang |
| author_sort | Patricia G. Wilson |
| collection | DOAJ |
| description | Fetal cells are widely considered a superior cell source for regenerative medicine; fetal cells show higher proliferative capacity and have undergone fewer replicative cycles that could generate spontaneous mutations. Fetal cells in amniotic fluid were among the first normal primary cells to be cultured ex vivo, but the undefined composition of amniotic fluid has hindered advance for regenerative applications. We first developed a highly efficient method to generate clonal populations by dilution of amniocentesis samples in media and direct plating without intervening refrigeration, centrifugation, or exposure of cells to the paracrine effects in mixed cell cultures. More than 40 clonal populations were recovered from 4 amniocentesis samples and representative clones were characterized by flow cytometry, conventional assays for differentiation potential, immunofluorescence imaging, and transcript analysis. The results revealed previously unreported diversity among stromal and epithelial cell types and identified unique cell types that could be lost or undetected in mixed cell populations. The differentiation potential of amniotic cells proved to be uncoupled from expression of definitive cell surface or cytoplasmic markers for stromal and epithelial cells. Evidence for diversity among stromal and epithelial cells in amniotic fluid bears on interpretations applied to molecular and functional tests of amniotic cell populations. |
| format | Article |
| id | doaj-art-fbc8dd7e5c5741bfaa51780258f6a97f |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-fbc8dd7e5c5741bfaa51780258f6a97f2025-02-03T06:42:14ZengWileyStem Cells International1687-966X1687-96782012-01-01201210.1155/2012/485950485950Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden DiversityPatricia G. Wilson0Lorna Devkota1Tiffany Payne2Laddie Crisp3Allison Winter4Zhan Wang5Wake Forest School of Medicine, Institute for Regenerative Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USAWake Forest University, Medical Center Blvd., Winston-Salem, NC 27157, USAWake Forest University, Medical Center Blvd., Winston-Salem, NC 27157, USAWake Forest School of Medicine, Institute for Regenerative Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USAWake Forest School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USAWake Forest School of Medicine, Institute for Regenerative Medicine, Medical Center Blvd., Winston-Salem, NC 27157, USAFetal cells are widely considered a superior cell source for regenerative medicine; fetal cells show higher proliferative capacity and have undergone fewer replicative cycles that could generate spontaneous mutations. Fetal cells in amniotic fluid were among the first normal primary cells to be cultured ex vivo, but the undefined composition of amniotic fluid has hindered advance for regenerative applications. We first developed a highly efficient method to generate clonal populations by dilution of amniocentesis samples in media and direct plating without intervening refrigeration, centrifugation, or exposure of cells to the paracrine effects in mixed cell cultures. More than 40 clonal populations were recovered from 4 amniocentesis samples and representative clones were characterized by flow cytometry, conventional assays for differentiation potential, immunofluorescence imaging, and transcript analysis. The results revealed previously unreported diversity among stromal and epithelial cell types and identified unique cell types that could be lost or undetected in mixed cell populations. The differentiation potential of amniotic cells proved to be uncoupled from expression of definitive cell surface or cytoplasmic markers for stromal and epithelial cells. Evidence for diversity among stromal and epithelial cells in amniotic fluid bears on interpretations applied to molecular and functional tests of amniotic cell populations.http://dx.doi.org/10.1155/2012/485950 |
| spellingShingle | Patricia G. Wilson Lorna Devkota Tiffany Payne Laddie Crisp Allison Winter Zhan Wang Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity Stem Cells International |
| title | Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity |
| title_full | Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity |
| title_fullStr | Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity |
| title_full_unstemmed | Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity |
| title_short | Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity |
| title_sort | clonal populations of amniotic cells by dilution and direct plating evidence for hidden diversity |
| url | http://dx.doi.org/10.1155/2012/485950 |
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