Tri-lineage Differentiation of hUC-MSCs

hUC-MSCs at various passages (P3, P6, P15) were seeded at a density of 2 × 10⁴/cm² in MSC culture medium. When cells reached 80% fusion, the medium was changed into adipogenic (MesenCult Adipogenic Differentiation Kit, Stem Cell Technologies), osteogenic (MesenCult Osteogenic Differentiation Kit, Stem Cell Technologies), or chondrogenic (MesenCult-ACF Chondrogenic Differentiation Kit, Stem Cell Technologies) differentiation medium. The differentiation medium was changed every 3 days as we described previously [6 (link), 20 (link)]. Twenty-one days later, the hUC-MSC-derived cells were stained by Oil Red S, Alizarin Red, or Alcian Blue staining and photographed with Nikon ElipseTi-U microscope (Nikon, Tokyo, Japan). The primer sequences for tri-lineage differentiation are available in Additional file 7: Table S1.

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Zhao Q., Zhang L., Wei Y., Yu H., Zou L., Huo J., Yang H., Song B., Wei T., Wu D., Zhang W., Zhang L., Liu D., Li Z., Chi Y., Han Z, & Han Z. (2019). Systematic comparison of hUC-MSCs at various passages reveals the variations of signatures and therapeutic effect on acute graft-versus-host disease. Stem Cell Research & Therapy, 10, 354.

Publication 2019

MesenCult Adipogenic Differentiation Kit MesenCult Osteogenic Differentiation Kit MesenCult-ACF Chondrogenic Differentiation Kit Nikon ElipseTi-U microscope

Adipogenic Alcian blue Cells Chondrogenic Microscope Osteogenic Primer Red s Stem cell

Corresponding Organization :

Other organizations : Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Kunming Medical University, First Affiliated Hospital of Kunming Medical University, Affiliated Hospital of Southwest Medical University, Soochow University, First Affiliated Hospital of Soochow University, Jinan University, Nankai University

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Variable analysis

independent variables

Cell passage (P3, P6, P15)

dependent variables

Adipogenic differentiation
Osteogenic differentiation
Chondrogenic differentiation

control variables

Cell seeding density (2 × 10^4/cm^2)
Culture medium (MSC culture medium)
Differentiation medium (Adipogenic, Osteogenic, Chondrogenic differentiation media)
Differentiation duration (21 days)

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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