Multifaceted characterization of the biological and transcriptomic signatures of natural killer cells derived from cord blood and placental blood

Abstract

Background Perinatal blood including umbilical cord blood and placental blood are splendid sources for allogeneic NK cell generation with high cytotoxicity of combating pathogenic microorganism and malignant tumor. Despite the generation of NK cells from the aforementioned perinatal blood, yet the systematical and detailed information of the biological and transcriptomic signatures of UC-NKs and P-NKs before large-scale clinical applications in disease remodeling is still largely obscure. Methods Herein, we took advantage of the “3IL”-based strategy for high-efficient generation of NK cells from umbilical cord blood and placental blood (UC-NKs and P-NKs), respectively. On the one hand, we conducted flow cytometry (FCM) assay and coculture to evaluate the subpopulations, cellular vitality and cytotoxic activity of the aforementioned NK cells. On the other hand, with the aid of RNA-SEQ and multiple bioinformatics analyses, we further dissected the potential diversities of UC-NKs and P-NKs from the perspectives of transcriptomes. Results On the basis of the “3IL” strategy, high-efficient NKs were generated from mononuclear cells (MNCs) in perinatal blood. P-NKs revealed comparable ex vivo expansion but preferable activation and cytotoxicity upon K562 cells over UC-NKs. Both of the two NKs showed diversity in cellular vitality and transcriptome including apoptotic cells, cell cycle, gene expression profiling and the accompanied multifaceted biological processes. Conclusions Our data revealed the multifaceted similarities and differences of UC-NKs and P-NKs both at the cellular and molecular levels. Our findings supply new references for allogeneic NK cell-based immunotherapy in regenerative medicine and will benefit the further exploration for illuminating the underlying mechanism as well.

Characterization of the biological and transcriptomic landscapes of bone marrow-derived mesenchymal stem cells in patients with multiple myeloma

“…CCK-8 assay was performed for cell proliferation assessment as we recently described with several modifications [ 8 , 25 ]. Briefly, HD-MSCs and MM-MSCs were seeded in 96-well plates at a density of 2.5 × 10 3 /well, and OD450 values were detected by using the cell counting kit-8 (CCK-8) (BOSTER, China) and the microplate reader (Bio-Rad, USA) under absorbance at 450 nm at the indicated time points (0 h, 24 h, 72 h, 120 h) according to the manufacturer’s instruction.…”

“…For apoptotic detection, MM-MSCs and HD-MSCs were turned to the Annexin V Apoptosis Detection Kit (Sigma-Aldrich, USA) as we previously reported [ 18 , 25 ]. According to the manufacturer’s instructions, a total number of 1 × 10 6 cells were washed with 1 × PBS (Gibco, USA) and resuspended in 100 µL 1 × Binding Buffer.…”

Decoding the multifaceted signatures and transcriptomic characteristics of stem cells derived from apical papilla and dental pulp of human supernumerary teeth

“…Finally, the indicated SCAP‐Ss and DPSCs were washed with 1×PBS twice and turned to Canto II flow cytometer (BD Biosciences) for detection. All data were analyzed with FlowJo 7.6 software, as we recently reported (Gao et al, 2022). The detailed information on the antibodies is listed in Supporting Information: Table S1.…”

“…All data were analyzed with FlowJo 7.6 software, as we recently reported (Gao et al, 2022). The detailed information on the antibodies is listed in Supporting Information: Table S1.…”

Overcoming the challenges in translational development of natural killer cell therapeutics: An opinion paper

“…Other studies in this regard are focused on developing efficient methods and designing biomaterials for activation, expansion and isolation of NKCs (85, 88-91). Gao et al recently classified the biological and transcriptomic signatures of cord blood and placenta-derived NKCs revealing the cellular/molecular level similarities and differences existing between the NK cell types (92). More of such studies are needed as they serve as database for cell-based immunotherapy and will be beneficial to understanding and categorizing the mechanism of action of different NKCs.…”