human whole bone marrow aspirates  (Lonza)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Identification of a novel, MSC-induced macrophage subtype via single-cell sequencing: implications for intervertebral disc degeneration therapy

doi: 10.3389/fcell.2023.1286011

Figure Lengend Snippet: Comparison of flow cytometry sorted and Magnetic Assisted Cell Sorting (MACS) isolated CD14 + /CD11B + cells. (A) Overview of analyses. Human bone marrow aspirates were collected and sorted via flow cytometry or MACS. (B–G) Unfractionated nucleated bone marrow cells (upper panels) or MACS sorted cells (lower panels) were stained with anti-CD14 (B,C) , anti-CD11B (D,E) , or co-stained with anti-CD11B and anti-CD14 antibodies (F,G) . Flow cytometry was then performed to detect single and double stained cells in each sample. Unstained nucleated bone marrow cells were used as a negative control (H) .

Article Snippet: Human whole bone marrow aspirates from female, non-smokers aged 18–35 years old were procured from Lonza Corporation.

Techniques: Comparison, Flow Cytometry, FACS, Isolation, Staining, Negative Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Identification of a novel, MSC-induced macrophage subtype via single-cell sequencing: implications for intervertebral disc degeneration therapy

doi: 10.3389/fcell.2023.1286011

Figure Lengend Snippet: MSC Isolation and Survival in Hypoxia. MSCs were isolated from human bone marrow aspirates based on plastic adherence of whole bone marrow or expression of the STRO3 antigen. (A) Images from the first 6 days of culture post isolation for both the whole population of MSCs and STRO3 + MSCs. (B) Flow cytometry analysis of whole bone marrow and STRO3 + MSCs. (C) STRO3 + MSCs were cultured in hypoxia as shown and immunostained for the MSC surface marker Cd90 and DAPI stained. (D) STRO3+ MSCs were cultured in hypoxia for 48 h and immunostaining for Cd90 and BNIP3 was performed.

Article Snippet: Human whole bone marrow aspirates from female, non-smokers aged 18–35 years old were procured from Lonza Corporation.

Techniques: Isolation, Expressing, Flow Cytometry, Cell Culture, Marker, Staining, Immunostaining

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Label-free hematology analysis using deep-ultraviolet microscopy

doi: 10.1073/pnas.2001404117

Figure Lengend Snippet: Wide-field unfixed and unstained pseudocolorized UV image of a spiculated bone marrow aspirate (A) and the corresponding white-light bright-field microscopy image after fixing and staining (B). The red arrowheads point to spicules present in the smear. (Scale bars: 200 µm.)

Article Snippet: Whole human bone marrow aspirate was collected from healthy donors (AllCells) and added to Na heparin anticoagulant solution.

Techniques: Microscopy, Staining

Journal: Respiratory Research

Article Title: Mesenchymal stem cells promote alveolar epithelial cell wound repair in vitro through distinct migratory and paracrine mechanisms

doi: 10.1186/1465-9921-14-9

Figure Lengend Snippet: Characterisation of hMSC and alveolar A549 cells. ( A ) Human type II alveolar epithelial cell line, A549 cells were positive for type II AEC marker pro SP-C. Phase contrast image is taken from an independent field to the pro SP-C and pro SP-C/DAPI field. ( B ) hMSCs isolated from human bone marrow aspirates were positive for mesenchymal stem cell markers CD44, STRO-1, CD90 and CD146; and negative for haematopoietic markers CD14 and CD19. ( C ) hMSC tri-lineage differentiation: osteogenesis, adipogenesis and chondrogenesis were confirmed by cytochemical/immunocytochemistry staining with Alizarin Red/anti-Osteocalcin, Oil Red O/anti-FABP-4 and Alcian Blue/anti-Aggrecan respectively. Scale bars, 100 μm.

Article Snippet: Briefly, human whole bone marrow aspirate (collected from iliac crest) (Lonza, USA) was seeded at a density of 10 5 mononuclear cells/cm 2 on 10 ng/ml Fibronectin-coated (Cat. No. F0895, Sigma) T-75 tissue culture flasks in 20 ml of DMEM (High glucose, 4.5g/L) supplemented with 5% foetal bovine serum (FBS), 1% L-glutamine, 1% non-essential amino acid (NEAA) and 1% PSA (Penicillin-Streptomycin-Amphotericin B) without any prior gradient centrifugation or immunoselection.

Techniques: Marker, Isolation, Immunocytochemistry, Staining