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Primary bone marrow cells co-cultured <t>with</t> <t>HS-5</t> marrow stromal cells. (A) Visualization of the culture in 1 μM Hoechst 33258 (blue) and 0.5 μM NucView (green, indicating activated caspase 3 in cells undergoing apoptosis. (B) Image analysis inverting the blue channel from part A to differentiate HS-5 cells (large diffuse; red arrow) from bone marrow cells (small, roundish, and intense).
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Primary bone marrow cells co-cultured with HS-5 marrow stromal cells. (A) Visualization of the culture in 1 μM Hoechst 33258 (blue) and 0.5 μM NucView (green, indicating activated caspase 3 in cells undergoing apoptosis. (B) Image analysis inverting the blue channel from part A to differentiate HS-5 cells (large diffuse; red arrow) from bone marrow cells (small, roundish, and intense).

Journal: bioRxiv

Article Title: Efficacy evaluation of glasedgib Sonic Hedgehog pathway inhibition with or without inotuzumab in B-ALL cells using a new co-culturing system model and a validated chemosensitivity assay

doi: 10.64898/2026.05.07.723573

Figure Lengend Snippet: Primary bone marrow cells co-cultured with HS-5 marrow stromal cells. (A) Visualization of the culture in 1 μM Hoechst 33258 (blue) and 0.5 μM NucView (green, indicating activated caspase 3 in cells undergoing apoptosis. (B) Image analysis inverting the blue channel from part A to differentiate HS-5 cells (large diffuse; red arrow) from bone marrow cells (small, roundish, and intense).

Article Snippet: Patients’ cells were cultured in vitro with support of a feeder cell layer HS-5 stromal cells (ATCC CRL-11882).

Techniques: Cell Culture

(A) Experimental design: primary B-ALL blasts were recovered from cryostorage and briefly allowed to recover in co-culture with HS-5 stromal support. Viable blasts were plated for chemosensitivity testing in co-culture and exposed to treatments for four days. Hematopoietic cells were mechanically isolated and assessed for viability using CellTiter-Glo. (B) Relative viability: dose-dependent responses to glasdegib are in seen within the hematopoietic compartment of co-cultured cells (Patients 1 and 2) but not in HS-5 stromal cells alone. Unique patients have differential responses to inotuzumab alone, but co-treatment sensitizes blasts to the effect of glasdegib.

Journal: bioRxiv

Article Title: Efficacy evaluation of glasedgib Sonic Hedgehog pathway inhibition with or without inotuzumab in B-ALL cells using a new co-culturing system model and a validated chemosensitivity assay

doi: 10.64898/2026.05.07.723573

Figure Lengend Snippet: (A) Experimental design: primary B-ALL blasts were recovered from cryostorage and briefly allowed to recover in co-culture with HS-5 stromal support. Viable blasts were plated for chemosensitivity testing in co-culture and exposed to treatments for four days. Hematopoietic cells were mechanically isolated and assessed for viability using CellTiter-Glo. (B) Relative viability: dose-dependent responses to glasdegib are in seen within the hematopoietic compartment of co-cultured cells (Patients 1 and 2) but not in HS-5 stromal cells alone. Unique patients have differential responses to inotuzumab alone, but co-treatment sensitizes blasts to the effect of glasdegib.

Article Snippet: Patients’ cells were cultured in vitro with support of a feeder cell layer HS-5 stromal cells (ATCC CRL-11882).

Techniques: Co-Culture Assay, Isolation, Cell Culture

The SHH gene expressions by four primary B-ALL patient samples in different culture conditions. Each panel represents a different patient sample, and each color corresponds to the SHH gene indicated in the legend. The x-axes represent the treatment conditions: negative control (Co DMSO), 20 μM glasdegib (Co Glas 20), and 20 μM glasdegib + 10 ng/mL inotuzumab (Co Glas 20 INO). The y-axes represent the expression of GLI1, GLI3, SMO , and PTCH1 after four days of HS-5 stromal cell co-culture. All expression values were represented by method with GAPDH as the internal control and treatment with DMSO as the fold-change calibrator.

Journal: bioRxiv

Article Title: Efficacy evaluation of glasedgib Sonic Hedgehog pathway inhibition with or without inotuzumab in B-ALL cells using a new co-culturing system model and a validated chemosensitivity assay

doi: 10.64898/2026.05.07.723573

Figure Lengend Snippet: The SHH gene expressions by four primary B-ALL patient samples in different culture conditions. Each panel represents a different patient sample, and each color corresponds to the SHH gene indicated in the legend. The x-axes represent the treatment conditions: negative control (Co DMSO), 20 μM glasdegib (Co Glas 20), and 20 μM glasdegib + 10 ng/mL inotuzumab (Co Glas 20 INO). The y-axes represent the expression of GLI1, GLI3, SMO , and PTCH1 after four days of HS-5 stromal cell co-culture. All expression values were represented by method with GAPDH as the internal control and treatment with DMSO as the fold-change calibrator.

Article Snippet: Patients’ cells were cultured in vitro with support of a feeder cell layer HS-5 stromal cells (ATCC CRL-11882).

Techniques: Negative Control, Expressing, Co-Culture Assay, Control