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Journal: bioRxiv

Article Title: S3-enriched kidney proximal nephrons from stem cells facilitate tubular injury modelling

doi: 10.64898/2026.01.29.702488

Figure Lengend Snippet: A. Quantification of the viability of D13 organoid progenitors following thaw using the Countess™ 3 FL Automated Cell Counter, comparing D13 progenitors cryopreserved in Cryo-SFM (light grey bars) and 5% DMSO (dark grey bars). Error bars indicate SEM from 3 independent thawed D13 cryovials. B-C . Brightfield ( B ) and confocal immunofluorescence ( C ) images of PT-EKO at D13+14 generated from fresh D13 monolayers (control) or thawed D13 monolayers that had been cryopreserved in Cryo-SFM or DMSO. Immunofluorescence ( C ) depicts PT (LTL; blue), podocytes (NPHS1; grey), loop of Henle TAL (SLC12A1; red), and nephron epithelium (EPCAM; green). Scale bars represent 200µm. D. qRT-PCR of nephron patterning genes ( MAFB ; podocytes, LRP2 ; PT, SLC12A1 ; loop of Henle TAL, and ECAD ; distal nephron) in D13+14 control PT-EKO (cyan bars), and PT-EKO generated from Cryo-SFM (yellow bars) and DMSO (magenta bars) cryopreserved D13 monolayers. Error bars indicate SEM from n = 3 biological replicates. Statistical significance was assessed using a one-way ANOVA with Tukey’s multiple comparisons test. Asterisks (**) denote two-tailed P value ≤ 0.01 for pairwise comparison.

Article Snippet: Cell pellets were resuspended in ice-cold Cryo-SFM freezing medium (Promo-Cell, Heidelberg, Germany, cat# C-29912) at a density of approximately 6 x10 6 cells/mL before transferring 1mL aliquots into cryovials (Corning, New York, USA).

Techniques: Immunofluorescence, Generated, Control, Quantitative RT-PCR, Two Tailed Test, Comparison

Journal: bioRxiv

Article Title: S3-enriched kidney proximal nephrons from stem cells facilitate tubular injury modelling

doi: 10.64898/2026.01.29.702488

Figure Lengend Snippet: A. Quantification of the viability of D13 organoid progenitors following thaw using the Countess™ 3 FL Automated Cell Counter, comparing D13 progenitors cryopreserved in Cryo-SFM (light grey bars) and 5% DMSO (dark grey bars). Error bars indicate SEM from 3 independent thawed D13 cryovials. B-C . Brightfield ( B ) and confocal immunofluorescence ( C ) images of PT-EKO at D13+14 generated from fresh D13 monolayers (control) or thawed D13 monolayers that had been cryopreserved in Cryo-SFM or DMSO. Immunofluorescence ( C ) depicts PT (LTL; blue), podocytes (NPHS1; grey), loop of Henle TAL (SLC12A1; red), and nephron epithelium (EPCAM; green). Scale bars represent 200µm. D. qRT-PCR of nephron patterning genes ( MAFB ; podocytes, LRP2 ; PT, SLC12A1 ; loop of Henle TAL, and ECAD ; distal nephron) in D13+14 control PT-EKO (cyan bars), and PT-EKO generated from Cryo-SFM (yellow bars) and DMSO (magenta bars) cryopreserved D13 monolayers. Error bars indicate SEM from n = 3 biological replicates. Statistical significance was assessed using a one-way ANOVA with Tukey’s multiple comparisons test. Asterisks (**) denote two-tailed P value ≤ 0.01 for pairwise comparison.

Article Snippet: PT cells for cryopreservation were dissociated with TryPLE (Gibco) as described above and cells were resuspended in ice-cold Cryo-SFM (Promo-Cell, Heidelberg, Germany) freezing medium, at a density of approximately 500,000 cells/mL, allowing 0.5mL per cryovial.

Techniques: Immunofluorescence, Generated, Control, Quantitative RT-PCR, Two Tailed Test, Comparison