Journal: Journal of Biological Engineering

Article Title: CD24 identifies nucleus pulposus progenitors/notochordal cells for disc regeneration

doi: 10.1186/s13036-018-0129-0

Figure Lengend Snippet: CD24 + NP cells decreased with the severity of disc degeneration. a - b Western blot analysis of CD24 expression in NP tissues with non, moderate and severe degeneration. c - d Immunohistochemical staining analysis of CD24 expression in NP tissues with non, moderate and severe degeneration. Scale bar represent 25 μm. **P < 0.01 , ***P < 0.001 . P -values were analyzed by one-way ANOVA

Article Snippet: After that, we incubated NP cells with a phycoerythrin (PE)-conjugated anti-rat CD24 antibody (130–106-254, Miltenyi) for 30 min at 4 °C in the dark.

Techniques: Western Blot, Expressing, Immunohistochemical staining, Staining

Journal: Journal of Biological Engineering

Article Title: CD24 identifies nucleus pulposus progenitors/notochordal cells for disc regeneration

doi: 10.1186/s13036-018-0129-0

Figure Lengend Snippet: CD24 + NP cells express a higher level of notochordal/immature NP cell marker. a - c Immunofluorescence staining analysis of brachyury, SHH and GLUT-1 in CD24 + , CD24 − and unsorted NP cells. d - e Western blot analysis of KRT8, brachyury, SHH and GLUT-1 expression in CD24 + , CD24 − and unsorted NP cells. *P < 0.05 , **P < 0.01 , ***P < 0.001 . P -values were analyzed by t one-way ANOVA. Scale bars represent 50 μm

Article Snippet: After that, we incubated NP cells with a phycoerythrin (PE)-conjugated anti-rat CD24 antibody (130–106-254, Miltenyi) for 30 min at 4 °C in the dark.

Techniques: Marker, Immunofluorescence, Staining, Western Blot, Expressing

Journal: Journal of Biological Engineering

Article Title: CD24 identifies nucleus pulposus progenitors/notochordal cells for disc regeneration

doi: 10.1186/s13036-018-0129-0

Figure Lengend Snippet: CD24 + NP cells exhibit a higher proliferation rate and higher osteogenic/adipogenic/chondrogenic differentiation ability than CD24 − and unsorted NP cells in vitro. a The proliferative activity of CD24 + , CD24 − and unsorted NP cells detected by CCK8. b - c Osteogenic differentiation of CD24 + , CD24 − and unsorted NP cells analysed by alkaline phosphatase ( b ) and alizarin red staining ( c ). d Adipogenic differentiation of CD24 + , CD24 − and unsorted NP cells analysed by oil red O staining. e Chondrogenic differentiation of CD24 + , CD24 − and unsorted NP cells analysed by alcian blue staining. **P < 0.01 , ***P < 0.001 . P -values were analyzed by one-way ANOVA

Article Snippet: After that, we incubated NP cells with a phycoerythrin (PE)-conjugated anti-rat CD24 antibody (130–106-254, Miltenyi) for 30 min at 4 °C in the dark.

Techniques: In Vitro, Activity Assay, Staining

Journal: Journal of Biological Engineering

Article Title: CD24 identifies nucleus pulposus progenitors/notochordal cells for disc regeneration

doi: 10.1186/s13036-018-0129-0

Figure Lengend Snippet: Osteogenic/adipogenic/chondrogenic differentiation related genes expression in CD24 + /CD24 − and unsorted NP cells after 3 weeks of differentiation induction. a - d Quantification analysis of BMP2 ( a ), RUNX2 ( b ), osteocalcin ( c ) and alkaline phosphate ( d ) expression in CD24 + , CD24 − and unsorted NP cells after 3 weeks of osteogenic differentiation induction. e - h Quantification analysis of adiponectin ( e ), PPAR-γ ( f ), C/EBPα ( g ) and lipoprotein ligase ( h ) expression in CD24 + , CD24 − and unsorted NP cells after 3 weeks of adipogenic differentiation induction. i - j Quantification analysis of COL II ( i ) and SOX-9 ( j ) expression in CD24 + , CD24 − and unsorted NP cells after 3 weeks of chondrogenic differentiation induction. **P < 0.01 , ***P < 0.001 . P -values were analyzed by one-way ANOVA

Article Snippet: After that, we incubated NP cells with a phycoerythrin (PE)-conjugated anti-rat CD24 antibody (130–106-254, Miltenyi) for 30 min at 4 °C in the dark.

Techniques: Expressing

Journal: Journal of Biological Engineering

Article Title: CD24 identifies nucleus pulposus progenitors/notochordal cells for disc regeneration

doi: 10.1186/s13036-018-0129-0

Figure Lengend Snippet: CD24 + NP cells transplantation ameliorates disc degeneration. a lateral X-ray images at 0 and 4 weeks after CD24 + and unsorted NP cells transplantation into the NP tissue punctured with needle. b - c Quantification of disc height at 0 and 4 weeks after CD24 + and unsorted NP cells transplantation into the NP tissue punctured with needle. d - e Midsagittal T2-weighted images at 0 and 4 weeks after CD24 + and unsorted NP cells transplantation into the NP tissue punctured with needle. f Histological analysis of intervertebral discs by Safranin O/Fast Green staining, immunohistochemical staining of collagen II and MMP13 in IVDs. Scale bars represent 100 μm. g - i Western blot analysis of collagen II and MMP13 in IVDs. *P < 0.05 , **P < 0.01 , ***P < 0.001 . P -values were analyzed by two-tailed t tests in ( c ) and one-way ANOVA in ( e - i )

Article Snippet: After that, we incubated NP cells with a phycoerythrin (PE)-conjugated anti-rat CD24 antibody (130–106-254, Miltenyi) for 30 min at 4 °C in the dark.

Techniques: Transplantation Assay, Staining, Immunohistochemical staining, Western Blot, Two Tailed Test

Journal: Journal of Biological Engineering

Article Title: CD24 identifies nucleus pulposus progenitors/notochordal cells for disc regeneration

doi: 10.1186/s13036-018-0129-0

Figure Lengend Snippet: HIF-1α pathway activation is crucial for CD24 + NP cells maintenance. a - b Western blot analysis of HIF-1α expression in CD24 + , CD24 − and unsorted NP cells. c Immunofluorescence staining analysis of CD24 in WT and NP-specific HIF-1α knock out mice, scale bars represent 100 μm. d - e Fluorescence activated cell sorting analysis of the percentage of CD24 + NP cells after Hif-1α or Vhl knock down. f Immunofluorescence staining analysis of CD24 in NP cells after Hif-1α or Vhl knock down, scale bars represent 25 μm. *P < 0.05 , ***P < 0.001 . P -values were analyzed by one-way ANOVA

Article Snippet: After that, we incubated NP cells with a phycoerythrin (PE)-conjugated anti-rat CD24 antibody (130–106-254, Miltenyi) for 30 min at 4 °C in the dark.

Techniques: Activation Assay, Western Blot, Expressing, Immunofluorescence, Staining, Knock-Out, Fluorescence, FACS, Knockdown

Journal: Journal of Biological Engineering

Article Title: CD24 identifies nucleus pulposus progenitors/notochordal cells for disc regeneration

doi: 10.1186/s13036-018-0129-0

Figure Lengend Snippet: HIF-1α-NOTCH1 pathway activation is essential for CD24 + NP cells maintenance. a - b Western blot analysis of JAGGED-1, NOTCH1 and HES-1 expression in CD24 + , CD24 − and unsorted NP cells. (c) Quantification analysis of JAGGED-1, NOTCH1 and HES-1 mRNA expression in CD24 + , CD24 − and unsorted NP cells. d - e Fluorescence activated cell sorting analysis of the percentage of CD24 + NP cells after Hif-1α or Vhl knock down with or without DAPT and JAGGED-1 stimulation. *P < 0.05 , **P < 0.01, ***P < 0.001 . P -values were analyzed by one-way ANOVA

Article Snippet: After that, we incubated NP cells with a phycoerythrin (PE)-conjugated anti-rat CD24 antibody (130–106-254, Miltenyi) for 30 min at 4 °C in the dark.

Techniques: Activation Assay, Western Blot, Expressing, Fluorescence, FACS, Knockdown

Journal: Journal of Biological Engineering

Article Title: CD24 identifies nucleus pulposus progenitors/notochordal cells for disc regeneration

doi: 10.1186/s13036-018-0129-0

Figure Lengend Snippet: The schematic graph reflects HIF-1α-NOTCH1 in the maintenance of CD24 + NP cells. HIF-1α-NOTCH1 pathway is essential for the maintenance of CD24 + NP cells, and which drives disc regeneration

Article Snippet: After that, we incubated NP cells with a phycoerythrin (PE)-conjugated anti-rat CD24 antibody (130–106-254, Miltenyi) for 30 min at 4 °C in the dark.

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: Lumican from DESC spheroids enhances odontoblastic differentiation of SCAPs

doi: 10.1186/s13287-025-04849-7

Figure Lengend Snippet: Characterization of SCAPs and DESCs. a Microscopic image of the apical buds from the lower incisors of 5-7-day-old C57BL/6 mice. b Primary mixed cell population. c Purified SCAPs exhibiting spindle-shaped morphology. d Multidirectional differentiation potential of SCAPs. Alcian Blue, Alizarin Red, and Oil Red O staining confirmed glycosaminoglycan deposition, mineralized nodule formation, and lipid droplet accumulation, indicative of chondrogenic, osteogenic, and adipogenic differentiation, respectively. e , f Flow cytometry analysis revealed CD24, CD133, and CD90 positivity, and CD45 negativity in SCAPs ( n = 3). The raw dot plots are presented in Supplementary Fig. S3. g Purified DESCs displaying cobblestone-like morphology. h Immunofluorescence imaging demonstrated positive K14 expression in DESCs (K14, green; DAPI, blue). i , j Flow cytometry analysis revealed CD29 and CD49f positivity, and CD90 negativity in DESCs ( n = 3). The raw dot plots are presented in Supplementary Fig. S3

Article Snippet: For SCAP analysis, cells were labeled with allophycocyanin (APC)-conjugated CD24 antibody (APC-65121, Proteintech, China, 1:300) as well as FITC-conjugated CD133 (11-1331-82, Thermo Fisher Scientific, USA, 1:500), CD90, and CD45 (MCD4501, Thermo Fisher Scientific, USA, 1:500) antibodies.

Techniques: Purification, Staining, Flow Cytometry, Immunofluorescence, Imaging, Expressing