PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions
The PC12 cell line is one of the most commonly used in neuroscience research, including studies on neurotoxicity, neuroprotection, neurosecretion, neuroinflammation, and synaptogenesis. Two types of this line are available in the ATCC collection: traditional PC12 cells grown in suspension and well-a...
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| Format: | Article |
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MDPI AG
2020-04-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/4/958 |
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| author | Benita Wiatrak Adriana Kubis-Kubiak Agnieszka Piwowar Ewa Barg |
| author_facet | Benita Wiatrak Adriana Kubis-Kubiak Agnieszka Piwowar Ewa Barg |
| author_sort | Benita Wiatrak |
| collection | DOAJ |
| description | The PC12 cell line is one of the most commonly used in neuroscience research, including studies on neurotoxicity, neuroprotection, neurosecretion, neuroinflammation, and synaptogenesis. Two types of this line are available in the ATCC collection: traditional PC12 cells grown in suspension and well-attached adherent phenotype. PC12 cells grown in suspension tend to aggregate and adhere poorly to non-coated surfaces. Therefore, it is necessary to modify the surface of culture vessels. This paper aims to characterise the use of two distinct variants of PC12 cells as well as describe their differentiation and neuronal outgrowth with diverse NGF concentrations (rat or human origin) on various surfaces. In our study, we evaluated cell morphology, neurite length, density and outgrowth (measured spectrofluorimetrically), and expression of neuronal biomarkers (doublecortin and NeuN). We found that the collagen coating was the most versatile method of surface modification for both cell lines. For adherent cells, the coating was definitely less important, and the poly-<span style="font-variant: small-caps;">d</span>-lysine surface was as good as collagen. We also demonstrated that the concentration of NGF is of great importance for the degree of differentiation of cells. For suspension cells, we achieved the best neuronal characteristics (length and density of neurites) after 14 days of incubation with 100 ng/mL NGF (change every 48 h), while for adherent cells after 3–5 days, after which they began to proliferate. In the PC12 cell line, doublecortin (DCX) expression in the cytoplasm and NeuN in the cell nucleus were found. In turn, in the PC12 Adh line, DCX was not expressed, and NeuN expression was located in the entire cell (both in the nucleus and cytoplasm). Only the traditional PC12 line grown in suspension after differentiation with NGF should be used for neurobiological studies, especially until the role of the NeuN protein, whose expression has also been noted in the cytoplasm of adherent cells, is well understood. |
| first_indexed | 2024-03-10T20:28:43Z |
| format | Article |
| id | doaj.art-119ab229b1c9457d85603e47e584ec89 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T20:28:43Z |
| publishDate | 2020-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-119ab229b1c9457d85603e47e584ec892023-11-19T21:31:44ZengMDPI AGCells2073-44092020-04-019495810.3390/cells9040958PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture ConditionsBenita Wiatrak0Adriana Kubis-Kubiak1Agnieszka Piwowar2Ewa Barg3Department of Basic Medical Sciences, Wroclaw Medical University, 50-556 Wroclaw, PolandDepartment of Toxicology, Wroclaw Medical University, 50-556 Wroclaw, PolandDepartment of Toxicology, Wroclaw Medical University, 50-556 Wroclaw, PolandDepartment of Basic Medical Sciences, Wroclaw Medical University, 50-556 Wroclaw, PolandThe PC12 cell line is one of the most commonly used in neuroscience research, including studies on neurotoxicity, neuroprotection, neurosecretion, neuroinflammation, and synaptogenesis. Two types of this line are available in the ATCC collection: traditional PC12 cells grown in suspension and well-attached adherent phenotype. PC12 cells grown in suspension tend to aggregate and adhere poorly to non-coated surfaces. Therefore, it is necessary to modify the surface of culture vessels. This paper aims to characterise the use of two distinct variants of PC12 cells as well as describe their differentiation and neuronal outgrowth with diverse NGF concentrations (rat or human origin) on various surfaces. In our study, we evaluated cell morphology, neurite length, density and outgrowth (measured spectrofluorimetrically), and expression of neuronal biomarkers (doublecortin and NeuN). We found that the collagen coating was the most versatile method of surface modification for both cell lines. For adherent cells, the coating was definitely less important, and the poly-<span style="font-variant: small-caps;">d</span>-lysine surface was as good as collagen. We also demonstrated that the concentration of NGF is of great importance for the degree of differentiation of cells. For suspension cells, we achieved the best neuronal characteristics (length and density of neurites) after 14 days of incubation with 100 ng/mL NGF (change every 48 h), while for adherent cells after 3–5 days, after which they began to proliferate. In the PC12 cell line, doublecortin (DCX) expression in the cytoplasm and NeuN in the cell nucleus were found. In turn, in the PC12 Adh line, DCX was not expressed, and NeuN expression was located in the entire cell (both in the nucleus and cytoplasm). Only the traditional PC12 line grown in suspension after differentiation with NGF should be used for neurobiological studies, especially until the role of the NeuN protein, whose expression has also been noted in the cytoplasm of adherent cells, is well understood.https://www.mdpi.com/2073-4409/9/4/958PC12NGFdifferentiationcoatingcollagenpolylysine |
| spellingShingle | Benita Wiatrak Adriana Kubis-Kubiak Agnieszka Piwowar Ewa Barg PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions Cells PC12 NGF differentiation coating collagen polylysine |
| title | PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions |
| title_full | PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions |
| title_fullStr | PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions |
| title_full_unstemmed | PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions |
| title_short | PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions |
| title_sort | pc12 cell line cell types coating of culture vessels differentiation and other culture conditions |
| topic | PC12 NGF differentiation coating collagen polylysine |
| url | https://www.mdpi.com/2073-4409/9/4/958 |
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