Journal: Journal of Assisted Reproduction and Genetics

Article Title: Rescue in vitro maturation using ovarian support cells of human oocytes from conventional stimulation cycles yields oocytes with improved nuclear maturation and transcriptomic resemblance to in vivo matured oocytes

doi: 10.1007/s10815-024-03143-4

Figure Lengend Snippet: Treatment with OSC-IVM improves maturation rate of human denuded oocytes compared to an IVM media-matched control. A Schematic of the experimental co-culture IVM approach. hiPSCs were differentiated using inducible transcription factor overexpression to form ovarian support cells (OSCs). Human oocytes were obtained from patients in the clinic after standard gonadotropin stimulation, and immature oocytes (GV and MI) identified after denuding were allocated between the experimental OSC-IVM condition (OSC-IVM) or the control IVM media condition (Media-IVM) for IVM co-culture. Oocyte maturation and health were assessed after 24–28 h IVM co-culture, and oocytes were frozen for further analyses. The figure was created with BioRender.com. B Representative image of co-culture containing immature human oocytes ( n = 3) and OSCs. Scale bar: 200 µm. Denuded GV oocytes are seen with surrounding OSCs in suspension culture. C Maturation rate of oocytes after 24–28 h IVM experiments, including oocyte co-culture with OSCs (OSC-IVM), or in media control (Media-IVM). n indicates the number of individual oocytes in each culture condition. Error bars indicate mean ± SEM. The p -value is derived from unpaired t -test comparing experimental OSC-IVM to control Media-IVM. Due to low numbers of retrieved oocytes per donor, each group contains oocytes from predominantly non-overlapping donor groups, and pairwise comparisons are not utilized

Article Snippet: All donated immature oocytes were maintained in preincubation LAG Medium (Medicult, Cooper Surgical) at 37 °C for 2–3 h after cumulus removal prior to introduction to in vitro maturation conditions for 24 to 28 h (either Media-IVM or OSC-IVM).

Techniques: Control, Co-Culture Assay, Over Expression, Suspension, Derivative Assay

Journal: Journal of Assisted Reproduction and Genetics

Article Title: Rescue in vitro maturation using ovarian support cells of human oocytes from conventional stimulation cycles yields oocytes with improved nuclear maturation and transcriptomic resemblance to in vivo matured oocytes

doi: 10.1007/s10815-024-03143-4

Figure Lengend Snippet: Cell culture media conditions

Article Snippet: All donated immature oocytes were maintained in preincubation LAG Medium (Medicult, Cooper Surgical) at 37 °C for 2–3 h after cumulus removal prior to introduction to in vitro maturation conditions for 24 to 28 h (either Media-IVM or OSC-IVM).

Techniques: Cell Culture, Recombinant

Journal: Journal of Assisted Reproduction and Genetics

Article Title: Rescue in vitro maturation using ovarian support cells of human oocytes from conventional stimulation cycles yields oocytes with improved nuclear maturation and transcriptomic resemblance to in vivo matured oocytes

doi: 10.1007/s10815-024-03143-4

Figure Lengend Snippet: MII oocytes treated with OSC-IVM are enriched for genes related to oocyte maturation and embryo development. A Graphical illustration of the strategy to identify transcriptomic profile enrichment during the progression from the germinal vesicle (GV) to MII oocyte. Differentially expressed genes (DEG) were identified by comparing GVs to MII oocytes. B Venn diagram illustrating the gene enrichment in successfully matured MII versus fail-to-mature GVs treated with OSC-IVM versus Media-IVM oocytes. Numbers display the total number of genes identified. Selected functional enrichment analysis terms are shown for each subgroup. See Supplementary Fig. for extended details. C Venn diagram illustrating the gene depletion in successfully matured MII versus fail-to-mature GVs treated with OSC-IVM versus Media-IVM oocytes. Numbers display the total number of genes identified. Selected functional enrichment analysis terms are shown for each subgroup. See Supplementary Fig. for extended details

Article Snippet: All donated immature oocytes were maintained in preincubation LAG Medium (Medicult, Cooper Surgical) at 37 °C for 2–3 h after cumulus removal prior to introduction to in vitro maturation conditions for 24 to 28 h (either Media-IVM or OSC-IVM).

Techniques: Functional Assay

Journal: Journal of Assisted Reproduction and Genetics

Article Title: Rescue in vitro maturation using ovarian support cells of human oocytes from conventional stimulation cycles yields oocytes with improved nuclear maturation and transcriptomic resemblance to in vivo matured oocytes

doi: 10.1007/s10815-024-03143-4

Figure Lengend Snippet: Transcriptomics analysis reveals that oocytes matured under the OSC-IVM condition are transcriptionally closer to IVF MII oocytes than those oocytes matured in Media-IVM, despite no changes in morphological features. A Total Oocyte Scores (TOS) generated from imaging analysis of MII oocytes after 24–28 h IVM experiments and IVF-MII oocytes. n indicates the number of individual MII oocytes analyzed. Median (dashed lines) and quartiles (dotted lines) are indicated. ANOVA indicated no significant ( p = 0.5274) difference between the means of each condition. B Quantification of the angle between the first polar body (PB1) and spindle apparatus, derived from oocyte fluorescent imaging analysis (See Supplementary Fig. ), of oocytes co-cultured with OSCs (OSC-IVM) or in media control (Media-IVM), and IVF-MII oocytes. n indicates the number of individual oocytes analyzed from each condition. Number and percentage (%) of MII oocytes with no spindle assembly observed are also indicated below the axis labels in the dashed box. Median (dashed line) and quartiles (dotted line) are indicated. ANOVA statistical analysis found no significant difference (ns, p = 0.1155) between the means of each condition. C Scatterplot projections of oocyte transcriptomes generated from the GV fail-to-mature Signature Score (X axis) and IVF MII Signature Score (Y axis). Symbols are color-coded based on the experimental condition (OSC-IVM, Media-IVM, IVF-MII), and symbol shapes represent oocyte maturation stages (GV, MI, and MII). Each symbol represents one oocyte. Histograms on top depict distribution of MII oocytes across the GV fail-to-mature Signature Score axis. Histograms on the right depict distribution of MII oocytes across the IVF MII Signature Score axis. Histograms are color-coded based on the experimental condition. n = 114 oocytes. D BoxPlot of distribution of MII oocytes across IVF MII Signature Score (left) and GV fail-to-mature Signature Score (right). ANOVA statistical analysis found a significant difference between the OSC-IVM-MII oocytes and Media-IVM MII oocytes (** p = 0.0003) and between the IVF-MII oocytes and Media-IVM oocytes (**** p < 0.0001), as well as between the OSC-IVM-MII oocytes and the IVF-MII oocytes (** p < 0.0026)

Article Snippet: All donated immature oocytes were maintained in preincubation LAG Medium (Medicult, Cooper Surgical) at 37 °C for 2–3 h after cumulus removal prior to introduction to in vitro maturation conditions for 24 to 28 h (either Media-IVM or OSC-IVM).

Techniques: Generated, Imaging, Derivative Assay, Cell Culture, Control

Journal: Journal of Assisted Reproduction and Genetics

Article Title: Rescue in vitro maturation using ovarian support cells of human oocytes from conventional stimulation cycles yields oocytes with improved nuclear maturation and transcriptomic resemblance to in vivo matured oocytes

doi: 10.1007/s10815-024-03143-4

Figure Lengend Snippet: Pathway enrichment analysis reveals similarities between MII oocytes rescued from OSC-IVM and IVF-MII oocytes. A Dotplot displaying gene enrichment among Media-IVM MII oocytes, OSC-IVM MII oocytes, and IVF-MII oocytes. The bottom panel shows enrichment of Gene Ontology (GO) terms and KEGG and REACTOME pathways (see an extended version in Supplementary Fig. ). GO, gene ontology; MF, molecular function; BP, biological process; CC, cellular component; KEGG, KEGG pathway; REAC, Reactome pathway. B Heatmap of Gene Set Enrichment Analysis (GSEA) hallmarks among Media-IVM MII oocytes, OSC-IVM MII oocytes, and IVF-MII oocytes. Heatmap represents row-normalized gene expression using a color gradients scale ranging from higher (red) to lower (blue) relative levels

Article Snippet: All donated immature oocytes were maintained in preincubation LAG Medium (Medicult, Cooper Surgical) at 37 °C for 2–3 h after cumulus removal prior to introduction to in vitro maturation conditions for 24 to 28 h (either Media-IVM or OSC-IVM).

Techniques: Expressing

Journal:

Article Title: In Vivo Quantitative Microvasculature Phenotype Imaging of Healthy and Malignant Tissues Using a Fiber-Optic Confocal Laser Microprobe 1

doi:

Figure Lengend Snippet: Histologic finding of normal pancreas and two orthotopic PDAC tumors with K-ras, Ink4a, and Arf mutations. (a) Hematoxylin and eosin staining of normal pancreas showing an islet of Langerhans (yellow arrow) and the deeply basophilic columnar acinar cells (red arrow). (b) A 2-week-old tumor derived from the NB587 cell line showing lymphocytic infiltrate (yellow arrow) and effacement of the acinar architecture by the invading ductal cells (red arrow). (c) A 2-week-old tumor derived from the NB743 cell line harboring the same mutations as NB587 and served here as an additional confirmation of phenotypes. Similar to (b), poorly formed glands were present in densely fibrotic stroma (yellow arrow) within the pancreatic substance. Original magnifications, x200. (d) IHC staining with anti-CD31 antibody for vessel diameter measurement validation. Blue bars were manually drawn by connecting the shortest point of an area enclosed by positive staining (in light brown). Here, a normal pancreas is shown as an example. Black scale bar, 20 µm. (e) IHC staining of PDAC shows tumor surface capillaries. (f) Intravital multichannel confocal microscopy on live PDAC revealed spatial relation between tumor surface capillaries (false-colored in red) and the GFP+ tumors in green. Yellow scale bar, 200 µm.

Article Snippet: Imaging Probe A fluorescent blood pool agent was used to image the microvasculature of different organs and murine orthotopic pancreatic ductal tumor (Angiosense 680 IVM; VisEn Medical, Woburn, MA).

Techniques: Staining, Derivative Assay, Immunohistochemistry, Confocal Microscopy

Journal:

Article Title: In Vivo Quantitative Microvasculature Phenotype Imaging of Healthy and Malignant Tissues Using a Fiber-Optic Confocal Laser Microprobe 1

doi:

Figure Lengend Snippet: Characterization of microvasculature of pancreatic ductal adenocarcinoma over time. (a) Tumor area from the same set of mice (7 in each, 21 total) was measured at weeks 1 and 2. (b) Both lines of PDAC tumors did not show a significant change in vessel diameter by week 1. However, by week 2, both had significantly larger-diameter vessels compared to the control. (c) Both lines of PDAC tumors showed significant increase in vessel density by week 1; these differences became more accentuated by week 2. (d) Volume fraction also became significantly greater for the tumors by week 1. (e) Fractal dimension did not change by week 1, but became significantly elevated for both lines of tumor. (f) Lacunarity remained unchanged during the 2-week period. (g) A 3D scatterplot showing an increase in vessel density, diameter, and fractal dimension secondary to orthotopic PDAC, represented by an upward shift in all axes from the control group (dashed arrow). The two tumor lines mixed into one cluster. Each point represented one measurement.

Article Snippet: Imaging Probe A fluorescent blood pool agent was used to image the microvasculature of different organs and murine orthotopic pancreatic ductal tumor (Angiosense 680 IVM; VisEn Medical, Woburn, MA).

Techniques:

Journal: Reproductive Medicine and Biology

Article Title: Oocyte collection and in vitro maturation after train transportation of human follicular fluid aspirated from resected non‐stimulated ovaries of patients with endometrial adenocarcinoma

doi: 10.1002/rmb2.12265

Figure Lengend Snippet: Techniques for transporting human follicular fluid. A, Appearance of the 300‐mL vacuum‐insulated bottle and the Falcon conical tube. B, Falcon conical tube in a 300‐mL vacuum‐insulated bottle filled with warm water (37°C). C, CellBox ® and Thermopack ® . D, Appearance of CulturePal ® inside CellBox ® . E, Dish with HFF and Sage IVM maturation medium is contained in CulturePal ® . White bag shown by red arrow is the CulturePal ® CO 2 generator. F, Schema of Thermopack ® and CulturePal ® in CellBox ®

Article Snippet: We washed the collected oocytes in Sage IVM washing medium (Cooper Surgical), transferred them to Sage IVM maturation medium (Cooper Surgical) with 75 IU of FSH and 75 IU of luteinizing hormone (HMG Injection Teizo; Asuka Pharmaceutical, Tokyo, Japan), and kept them in an incubator (5% CO , 5% O ; 37°C) for 24 hours.

Techniques:

Journal: Reproductive Medicine and Biology

Article Title: Oocyte collection and in vitro maturation after train transportation of human follicular fluid aspirated from resected non‐stimulated ovaries of patients with endometrial adenocarcinoma

doi: 10.1002/rmb2.12265

Figure Lengend Snippet: Collected oocytes from resected ovaries and immunofluorescent staining. A, Collected immature oocytes before in vitro maturation in patient 8. B, An oocyte in meiosis II showing the first polar body after in vitro maturation in patient 8. C, Immunofluorescent staining of a mature oocyte in patient 5. D, Immunofluorescent staining of a mature oocyte in patient 13. The spindle is enclosed in a square box. E, Enlarged view of the spindle corresponding to the boxed area in (D). White scale bar indicates 50 µm. Red scale bar indicates 25 µm. Green, microtubules; blue, DAPI

Article Snippet: We washed the collected oocytes in Sage IVM washing medium (Cooper Surgical), transferred them to Sage IVM maturation medium (Cooper Surgical) with 75 IU of FSH and 75 IU of luteinizing hormone (HMG Injection Teizo; Asuka Pharmaceutical, Tokyo, Japan), and kept them in an incubator (5% CO , 5% O ; 37°C) for 24 hours.

Techniques: Staining, In Vitro

Journal: Reproductive Medicine and Biology

Article Title: Oocyte collection and in vitro maturation after train transportation of human follicular fluid aspirated from resected non‐stimulated ovaries of patients with endometrial adenocarcinoma

doi: 10.1002/rmb2.12265

Figure Lengend Snippet: Comparison of maturation rates in the transportation and non‐transportation groups. Non‐Transport, non‐transportation group; Bottle, bottle group; Box, box group. The bars indicate the percentages of oocytes that reached mature metaphase II as a result of in vitro maturation. Two‐sided Fisher's exact test was used for statistical analysis

Article Snippet: We washed the collected oocytes in Sage IVM washing medium (Cooper Surgical), transferred them to Sage IVM maturation medium (Cooper Surgical) with 75 IU of FSH and 75 IU of luteinizing hormone (HMG Injection Teizo; Asuka Pharmaceutical, Tokyo, Japan), and kept them in an incubator (5% CO , 5% O ; 37°C) for 24 hours.

Techniques: Comparison, In Vitro