4.4 Article

Acoustic waves improves retroviral transduction in human retinal stem cells

Journal

JOURNAL OF THE CHINESE MEDICAL ASSOCIATION
Volume 81, Issue 9, Pages 830-836

Publisher

ELSEVIER TAIWAN
DOI: 10.1016/j.jcma.2018.05.002

Keywords

Gene transfection; Photoreceptor; Retinal stem cells; Retrovirus; Ultrasound standing waves

Funding

  1. Taipei Veterans General Hospital [V95E2-007, V95B2-013]
  2. Joint Projects of UTVGH [VGHUST95-P1-05]
  3. National Science Council [NSC-94, NSC-95, NSC 101-2314-B-341 -003 -MY3]
  4. Yen Tjing Ling Medical Foundation
  5. National Yang-Ming University
  6. National Taiwan University Seed Fund

Ask authors/readers for more resources

Backgrounds: The plasticity of retinal stem cells (RSCs), a type of cells that can differentiate into neuron cells and photoreceptor cells, endows them with potential therapeutic properties that can be applied to regenerative medicine. Gene modification of these stem cells before trans differentiation and transplantation enhances their survival and increases their therapeutic function. The different ways to effectively deliver gene into RSCs are still discussed. This study aimed to use the acoustic waves to improve the efficacy of gene delivery for RSCs. Methods: RSCs were obtained from non-fetal human ocular pigmented ciliary margin tissues. The enhanced green fluorescent protein-encoded murine stem cell retroviruses (MSCV) were prepared and used to infect RSCs. Glass chambers containing RSCs, retroviruses, and various concentrations of polybrene (0, 0.8, 2, 4 and 8 mu g/mL) were exposed under 20 or 25 Vp-p ultrasonic standing wave fields (USWF) for 5 min. The percentage of green fluorescent protein positive cells in each sample was calculated and compared to test the efficacy of gene transduction. Results: Our results showed that the efficiency of gene transduction by MSCV infection was enhanced following the concentration of polybrene and the energy of USWF. The percentage of green fluorescent protein positive cells was significantly higher in chambers that contained 8 1.1 mu g/mL of polybrene and was exposed to 20Vp-p of USWF for 5 min. In addition, the percentage increased in chambers contained 2, 4 and 8 mu g/mL of polybrene when they were exposed to 25Vp-p of USWF. Comparing to those did not treated with ultrasound, the efficiency of retroviral transduction to RSCs increased 4-fold after exposed to USWF for 5 min. Conclusion: We demonstrated the ability of ultrasound standing waves to improve retroviral transduction into RSCs. We believe that this may be applied to the experimental designs of future studies and may have possible therapeutic uses. Copyright (C) 2018, the Chinese Medical Association. Published by Elsevier Taiwan LLC.

Authors

I am an author on this paper
Click your name to claim this paper and add it to your profile.

Reviews

Primary Rating

4.4
Not enough ratings

Secondary Ratings

Novelty
-
Significance
-
Scientific rigor
-
Rate this paper

Recommended

No Data Available
No Data Available