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Research ArticleOpen Access

Efficient Expansion of Human Umbilical Cord Blood- Derived NK Cells Ex Vivo without Requiring Feeder Layers

Volume 8 - Issue 5

Chu Thi Thao1#, Do Thi Hoai Thu1,2#, Bui Viet Anh1, Truong Linh Huyen1, Nguyen Van Phong1, Nguyen Thanh Liem1 and Hoang Thi My Nhung*1,2

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    • 1Vinmec Research Institute of Stem cells and Gene Technology, Vietnam
    • 2VNU University of Science, Vietnam
    • #These two authors contributes equally to this manuscript.

    *Corresponding author: Hoang Thi My Nhung, Vinmec Research Institute of Stem cells and Gene Technology, VNU University of Science, Hanoi, Vietnam

Received: September 03, 2018;   Published: September 10, 2018

DOI: 10.26717/BJSTR.2018.08.001711

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Abstract

Introduction: Natural killer (NK) cells are central components of the innate immunity. They have ability to kill a wide range of cancer cells and are a promising tool for both autologous and allogeneic immune enhancement therapy in cancer treatment. Actually, NK cells can be derived from multiple sources such as: peripheral blood, cord blood… Among these, cord blood (CB) is known as an ideal for NK cells expansion because it occupies a higher percentage of NK cells compare to the peripheral blood and is a rich source of hematopoietic stem cell as well as progenitor immune cells. The objectives of this study were to invent a reasonable approach for expanding a relevant number of NK cells from human cord blood to clinical application. Methods: At the initial step, cord blood was collected from the patients and then the mononuclear cells were obtained by density centrifugation with Ficoll.

After that, these cells were cultured in 2 stages: stimulation and expansion with inactivated autologous plasma and BINKIT expansion kit contains several kinds of growth factors which are specified for NK cells. The immunophenotype of NK cells was analyzed every 2 days through %CD56+CD3- by Flow cytometry technique. The ex-vivo activation and expansion of NK cells was performed in GMP grade -clean room for about 3 weeks. Results: After culturing periods, the average NK cell count post-expansion was 2040.6 ± 1463.5 x10^6 and the average cell fold expansion was 814.4 ± 560.9, the expanded cells presented 90.6% ± 8.9% purify of CD56+CD3-, the percentage of CD56 bright CD3- cells was 84.2% ± 14.4%.Conclusion: Human cord blood has a significant potential to expand NK cells to relevant number for clinical use and our method are suitable for getting a relevant number and quality of NK cells to the clinical use.

Keywords: Natural Killer Cells; Cord Blood; Autologous Plasma; Growth Factors; Feeder Layer; Cytotoxic T Cells; Tumour; NK Cocktail; Ficoll Solution; Immunoglobulin

Abbreviations: NK: Natural Killer; UCB: Umbilical Cord Blood; CTL: Cytotoxic T Cells; IFN-γ: Interferon Gamma; IL: Interleukin; ILCs: Innate Lymphoid Cells; KIRs: Killer Cell Immunoglobulin (Ig) like Receptors; MHC: Major Histocompatibility Complex; MNC: Mononuclear Cell; PBMCs: Peripheral Blood Mononuclear Cells

Abstract | Introduction | Materials and Methods | Results | Discussion | Conclusion | Competing Interests | Acknowledgement | References |