An effective and efficient method of transfecting primary human chondrocytes in suspension.

Autor: Makki MS; Department of Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH 44272, United States., Akhtar N; Department of Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH 44272, United States., Haqqi TM; Department of Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH 44272, United States. Electronic address: thaqqi@neomed.edu.
Jazyk: angličtina
Zdroj: Analytical biochemistry [Anal Biochem] 2017 Jun 01; Vol. 526, pp. 29-32. Date of Electronic Publication: 2017 Mar 14.
DOI: 10.1016/j.ab.2017.03.009
Abstrakt: Human chondrocytes accumulate an ECM-rich matrix by secreting matrix macromolecules during monolayer culture, which makes them difficult to transfect efficiently. Here we report a non-viral based protocol to transfect the primary human chondrocytes with high efficiency in suspension. Chondrocyte cultures were digested using Pronase and Collagenase and transfected in suspension. Transfection efficiencies of more than 80% were achieved routinely using the protocol described. The viability of siRNA transfected or un-transfected chondrocytes was not affected and resulted in 80-90% knockdown of the target mRNA levels. This protocol may be useful in gene knockdown, and ectopic overexpression studies in chondrocytes.
(Copyright © 2017 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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