Journal: bioRxiv

Article Title: A Mesenchymal Cell Niche in Skin for Acute Myeloid Leukemia

doi: 10.1101/2022.05.20.491183

Figure Lengend Snippet: ( A ) A representative FACS profile showing FACS sorting/analysis of the Ebf2 + and Ebf2 − cells in dorsal skin. The cells were first gated within non-hematopoietic (CD45 − TER119 − ) and non-endothelial (CD31 − ) live (PI − ) stromal cells. Then, these cells lacking expression of CD44 were further analyzed for their expression of SCA1, PDGFRa/CD140a (PαS) and CD51. The numbers in the panel are the mean frequencies. ( B-C ) The Ebf2 + cell frequency within total PI − ( B ) or PI − CD45 − TER119 − CD31 − stromal cells ( C ) in dorsal skin. ( D ) The fractions of PαS cells within the Ebf2 + and Ebf2 − stromal cells. Each dot in B-D represents data from a single mouse in 3-6 experiments with the horizontal line as a mean value. ( E ) CFU-Fs in the Ebf2 + and Ebf2 − stromal cells. ( F ) CFU-Fs were exclusively found in the Ebf2 + SCA1 + cell fraction. Data in E-F are from 3 independent experiments and each dot represents replicate assays from 2-3 mice in each experiment. The horizontal line represents mean value. Wilcoxon matched-signed pair rank test was used for statistical analysis. (G-J) Single-cell analysis of CFU-Fs and lineage differentiation from the FACS-sorted Ebf2 + ( G-H ) and Ebf2 − PαS ( I-J ) stromal cells. The CFU-F frequencies ( G, I ) were determined by limiting dilution at a density of 1, 2, 5, 10 cells per well in a 96-well plate and the frequency of the single cells with bi-lineage plasticity ( H, J ) were assessed by multilineage differentiation potential of single CFU-Fs derived from the cells. ( K ) Representative images of the osteogenic and adipogenic differentiation from single CFU-F clones derived from Ebf2 + and Ebf2 − PαS cells. ( L ) Population-doubling time (PDT) of randomly selected CFU-Fs derived from single Ebf2 + and Ebf2 − PαS cells. Each line represents the growth kinetics of a single clone. See also in Figure S1.

Article Snippet: For osteogenic differentiation, cells were cultured with complete osteogenic medium mixed by human/mouse StemXVivo osteogenic / adipogenic base medium (CCM007, R&D Systems) and mouse StemXVivo ostegenic supplement (CCM009; R&D Systems) under normoxic condition for 14-21 days.

Techniques: Expressing, Single-cell Analysis, Derivative Assay, Clone Assay

Journal: Scientific Reports

Article Title: Enhanced cryopreservation of MSCs in microfluidic bioreactor by regulated shear flow

doi: 10.1038/srep35416

Figure Lengend Snippet: Multilineage differentiation of shear induced cryopreserved MSCs showing differentiation potential towards both osteogenic and adipogenic lineages. Adipogenic oil droplet formation and osteogenic calcium nodule formation was determined by Oil Red O and alizarin red assays respectively.

Article Snippet: Ficoll hypaque solution (Hisep LSM 1073), adipogenic differentiation media, Triton X-100, formaldehyde, MTT assay, adipogenic and osteogenic differentiation media were purchased from Hi-Media labs, India.

Techniques: Shear