An innovative cell selection approach in developing human cells overexpressing aspartyl/asparaginyl β-hydroxylase

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

Bakhtiari Hadi; PALIZBAN ABBASALI; KHANAHMAD HOSSEIN; MOFID MOHAMMAD REZA

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: RESEARCH IN PHARMACEUTICAL SCIENCES (RPS) Year:2020 | Volume:15 | Issue:3 Page(s): 291-299

Download Full-Text

View:

144

Download:

Cites:

Information Journal Paper

Title

An innovative cell selection approach in developing human cells overexpressing aspartyl/asparaginyl β-hydroxylase

Author(s)

Bakhtiari Hadi | PALIZBAN ABBASALI | KHANAHMAD HOSSEIN | MOFID MOHAMMAD REZA | Issue Writer Certificate

Keywords

Cancer

Aspartyl/asparaginyl β

-hydroxylase

Human embryonic kidney (HEK293T) cell line

Transfection

Overexpression

Abstract

Background and purpose: Aspartyl/asparaginyl β;-hydroxylase (ASPH) is abundantly expressed in malignant neoplastic cells. The establishment of a human cell line overexpressing ASPH could provide the native-like recombinant protein needed for developing theranostic probes. In the process of Transfection, the obtained cells normally contain a range of cells expressing the different levels of the target of interest. In this paper, we report on our simple innovative approach in the selection of best-transfected cells with the highest expression of ASPH using subclone selection, quantitative real-time polymerase chain reaction, and gradual increment of hygromycin concentration. Experimental approach: To achieve this goal, human embryonic kidney (HEK 293T) cells were transfected with an ASPH-bearing pcDNA3. 1/Hygro(+) vector. During antibiotic selection, single accumulations of the resistant cells were separately cultured and the ASPH mRNA levels of each flask were evaluated. The best subclones were treated with a gradually increasing amount of hygromycin. The ASPH protein expression of the obtained cells was finally evaluated using flow cytometry and immunocytochemistry. Findings / Results: The results showed that different selected subclones expressed different levels of ASPH. Furthermore, the gradual increment of hygromycin (up to 400mg/mL) improved the expression of ASPH. The best relative fold change in mRNA levels was 57. 59 ± 4. 11. Approximately 90. 2% of HEKASPH cells overexpressed ASPH on their surface. Conclusion and implications: The experiments indicated that we have successfully constructed and evaluated a recombinant human cell line overexpressing ASPH on the surface. Moreover, our innovative selection approach provided an effective procedure for enriching highly expressing recombinant cells.

Multimedia

No record.

Cites

No record.

References

No record.

Cite

APA: Copy

Bakhtiari, Hadi, PALIZBAN, ABBASALI, KHANAHMAD, HOSSEIN, & MOFID, MOHAMMAD REZA. (2020). An innovative cell selection approach in developing human cells overexpressing aspartyl/asparaginyl β-hydroxylase. RESEARCH IN PHARMACEUTICAL SCIENCES (RPS), 15(3), 291-299. SID. https://sid.ir/paper/318512/en

Vancouver: Copy

Bakhtiari Hadi, PALIZBAN ABBASALI, KHANAHMAD HOSSEIN, MOFID MOHAMMAD REZA. An innovative cell selection approach in developing human cells overexpressing aspartyl/asparaginyl β-hydroxylase. RESEARCH IN PHARMACEUTICAL SCIENCES (RPS)[Internet]. 2020;15(3):291-299. Available from: https://sid.ir/paper/318512/en

IEEE: Copy

Hadi Bakhtiari, ABBASALI PALIZBAN, HOSSEIN KHANAHMAD, and MOHAMMAD REZA MOFID, “An innovative cell selection approach in developing human cells overexpressing aspartyl/asparaginyl β-hydroxylase,” RESEARCH IN PHARMACEUTICAL SCIENCES (RPS), vol. 15, no. 3, pp. 291–299, 2020, [Online]. Available: https://sid.ir/paper/318512/en

Related Journal Papers

No record.

Related Seminar Papers

No record.

Related Plans

No record.

Recommended Workshops