Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

CD19/CD22 bivalent CAR T cells in children, adolescents and young adults with B-ALL: final phase 1 trial results.

Journal for immunotherapy of cancer·2026
Same author

Distinct in vivo dynamics of donor-derived stem cell memory CAR T cells post-allogeneic HSCT relapse.

Cell·2026
Same author

Bringing in vivo CAR to the clinic: Promise, pitfalls, and questions to ask.

Molecular therapy : the journal of the American Society of Gene Therapy·2026
Same author

A multicentre analysis of efficacy, safety and molecular response correlates of fostamatinib in warm autoimmune haemolytic anaemia and Evans syndrome.

British journal of haematology·2026
Same author

Alpha-gal sensitization and allergic blood transfusion reactions: a scoping review.

Journal of translational medicine·2026
Same author

Ten-year experience of CD22 CAR T cells for children and young adults with B-cell acute lymphoblastic leukemia.

Blood advances·2025

Related Experiment Video

Updated: Oct 12, 2025

An Efficient In Vitro Transposition Method by a Transcriptionally Regulated Sleeping Beauty System Packaged into an Integration Defective Lentiviral Vector
10:13

An Efficient In Vitro Transposition Method by a Transcriptionally Regulated Sleeping Beauty System Packaged into an Integration Defective Lentiviral Vector

Published on: January 12, 2018

9.1K

High efficiency closed-system gene transfer using automated spinoculation.

Victoria Ann Remley1, Jianjian Jin1, Sarmila Sarkar1

  • 1Center for Cellular Engineering, Department of Transfusion Medicine, NIH Clinical Center, Bethesda, USA.

Journal of Translational Medicine
|November 25, 2021
PubMed
Summary
This summary is machine-generated.

Spinoculation significantly enhances lentiviral gene transfer into lymphocytes for cell therapies. An automated closed-system method (Sepax spinoculation) improves efficiency and reduces contamination risks compared to traditional methods.

Keywords:
CAR T-cellGene transferSepaxSpinoculation

More Related Videos

Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor
07:28

Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

Published on: February 1, 2015

20.0K
Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

16.7K

Related Experiment Videos

Last Updated: Oct 12, 2025

An Efficient In Vitro Transposition Method by a Transcriptionally Regulated Sleeping Beauty System Packaged into an Integration Defective Lentiviral Vector
10:13

An Efficient In Vitro Transposition Method by a Transcriptionally Regulated Sleeping Beauty System Packaged into an Integration Defective Lentiviral Vector

Published on: January 12, 2018

9.1K
Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor
07:28

Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

Published on: February 1, 2015

20.0K
Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

16.7K

Area of Science:

  • Cellular Therapy
  • Gene Transfer Technology
  • Bioprocessing

Background:

  • Lentiviral vectors are crucial for cellular therapies, with spinoculation being an effective gene transfer method for lymphocytes and hematopoietic progenitor cells.
  • Current Good Manufacturing Practice (cGMP)-compliant production necessitates closed-system methods to minimize contamination and manual handling.

Purpose of the Study:

  • To develop and compare closed-system spinoculation methods for lentiviral gene transfer in lymphocytes.
  • To evaluate an automated closed system (Sepax spinoculation) for improved efficiency and reduced contamination in cell therapy production.

Main Methods:

  • Compared Sepax spinoculation, bag spinoculation, and static bag transduction for lentiviral gene transfer in apheresis-derived lymphocytes.
  • Assessed transduction efficiency of various Chimeric Antigen Receptor (CAR) lentiviral vectors (CD19/CD22, FGFR4, CD22).
  • Evaluated functional capacity (killing assays, cytokine secretion) and gene expression of transduced CAR T-cells.

Main Results:

  • Spinoculation significantly enhanced gene transfer efficiency compared to static methods.
  • Sepax spinoculation (1-h spin time) demonstrated comparable efficiency to bag spinoculation and superior results to static transduction (83.4% vs. 72.8% vs. 35.7%).
  • The methods were consistent across different CAR vectors and cell sources (healthy donor vs. patient), and did not impair CAR T-cell function or essential gene expression.

Conclusions:

  • Closed-system bag spinoculation offers more efficient lymphocyte gene transfer than standard methods, applicable to both retroviral and lentiviral vectors.
  • Sepax spinoculation provides an automated, closed-system approach that reduces hands-on time and contamination risks.
  • This automated method is a feasible advancement for gene transfer in lymphocytes and potentially other cell types for cell therapy production.