Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture Plate
Cervical cancer recurs in 90% cases and linked to cancer stem cells that able to self-renew and responsible for recurrence, metastasis, and mortality of cancer. Isolation and identification of cancer stem cells using serum-free medium needs expensive growth factors and consume time. This study try t...
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doaj-aa86d20b6e3042d19ab998cbcc69153c2020-11-24T22:28:19ZengUniversity of BrawijayaJournal of Tropical Life Science2087-55172527-43762016-11-016319620410.11594/jtls.06.03.11189Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture PlatePutu Juniartha0Muhammad Rasjad Indra1Hidayat Sujuti2Diana Lyrawati3Tatit Nurseta4Brawijaya UniversityBrawijaya UniversityBrawijaya UniversityBrawijaya UniversityBrawijaya UniversityCervical cancer recurs in 90% cases and linked to cancer stem cells that able to self-renew and responsible for recurrence, metastasis, and mortality of cancer. Isolation and identification of cancer stem cells using serum-free medium needs expensive growth factors and consume time. This study try to grow tumor sphere using culture plate coated with 1% agarose as an efficient and economical alternative to non-adhesive culture plate. HeLa cell line was grew in culture plate coated with 1% agarose and non-adhesive culture plate using similar medium and culture condition. Tumor spheres morphology was observed and the colonies were counted in 7 days followed by single cell assay. Tumor spheres then counted for CD133+, CD34+, and Sox2 expression using flowcytometry. Culture plate coated with 1% agarose can be used as an economic and efficient alternative to culture tumor sphere. Using culture plate coated with 1% agarose, the tumor spheres formed in 7 days with similar morphology to non-adhesive culture plate. Tumorsphere had three dimensional – sphere shape that tightly attached, colonized, and overlapped. The tumor sphere colony counts of two plates were statistically have no significant difference (p=0,667). Single cell assay of a tumor sphere shows that it can grow new tumor spheres with similar morphology. The tumor sphere from culture plate coated with 1% agarose express CD133+ and CD34+ as much as 8.78% ± 2.14 and Sox2 as much as 35.30% ± 23.82 whereas tumor sphere from non-adhesive culture plate express CD133+ and CD34+ as much as 62.36% ± 1.06 and Sox2 as much as 98.86% ± 0.56 (p = 0000).http://jtrolis.ub.ac.id/index.php/jtrolis/article/view/596Cancer stem cell, tumorsphere, agarose, non adhesive culture plate, Sox2. |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Putu Juniartha Muhammad Rasjad Indra Hidayat Sujuti Diana Lyrawati Tatit Nurseta |
spellingShingle |
Putu Juniartha Muhammad Rasjad Indra Hidayat Sujuti Diana Lyrawati Tatit Nurseta Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture Plate Journal of Tropical Life Science Cancer stem cell, tumorsphere, agarose, non adhesive culture plate, Sox2. |
author_facet |
Putu Juniartha Muhammad Rasjad Indra Hidayat Sujuti Diana Lyrawati Tatit Nurseta |
author_sort |
Putu Juniartha |
title |
Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture Plate |
title_short |
Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture Plate |
title_full |
Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture Plate |
title_fullStr |
Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture Plate |
title_full_unstemmed |
Agarose Coated Culture Plate in Tumorsphere Culture of Cervical Cancer Cell Line HeLa: an Alternative to Non Adhesive Culture Plate |
title_sort |
agarose coated culture plate in tumorsphere culture of cervical cancer cell line hela: an alternative to non adhesive culture plate |
publisher |
University of Brawijaya |
series |
Journal of Tropical Life Science |
issn |
2087-5517 2527-4376 |
publishDate |
2016-11-01 |
description |
Cervical cancer recurs in 90% cases and linked to cancer stem cells that able to self-renew and responsible for recurrence, metastasis, and mortality of cancer. Isolation and identification of cancer stem cells using serum-free medium needs expensive growth factors and consume time. This study try to grow tumor sphere using culture plate coated with 1% agarose as an efficient and economical alternative to non-adhesive culture plate. HeLa cell line was grew in culture plate coated with 1% agarose and non-adhesive culture plate using similar medium and culture condition. Tumor spheres morphology was observed and the colonies were counted in 7 days followed by single cell assay. Tumor spheres then counted for CD133+, CD34+, and Sox2 expression using flowcytometry. Culture plate coated with 1% agarose can be used as an economic and efficient alternative to culture tumor sphere. Using culture plate coated with 1% agarose, the tumor spheres formed in 7 days with similar morphology to non-adhesive culture plate. Tumorsphere had three dimensional – sphere shape that tightly attached, colonized, and overlapped. The tumor sphere colony counts of two plates were statistically have no significant difference (p=0,667). Single cell assay of a tumor sphere shows that it can grow new tumor spheres with similar morphology. The tumor sphere from culture plate coated with 1% agarose express CD133+ and CD34+ as much as 8.78% ± 2.14 and Sox2 as much as 35.30% ± 23.82 whereas tumor sphere from non-adhesive culture plate express CD133+ and CD34+ as much as 62.36% ± 1.06 and Sox2 as much as 98.86% ± 0.56 (p = 0000). |
topic |
Cancer stem cell, tumorsphere, agarose, non adhesive culture plate, Sox2. |
url |
http://jtrolis.ub.ac.id/index.php/jtrolis/article/view/596 |
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