Generation of two human induced pluripotent stem cell lines derived from myoblasts (MDCi014-A) and from peripheral blood mononuclear cells (MDCi014-B) from the same donor

Metzler, Eric; Telugu, Narasimha; Diecke, Sebastian; Spuler, Simone; Escobar, Helena

doi:10.1016/j.scr.2020.101998

Cited by 7 publications

(8 citation statements)

References 4 publications

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“…As previously noted in the literature, naïve hPSCs derived under 5iLA conditions exhibit low (38) to generally positive (39) levels of TRA-1-60 expression on the cell surface, while primed hPSCs display heterogeneous yet generally positive expression of this mark (fig. S3), quite similar to previous studies (37,(40)(41)(42)(43)(44)(45)(46)(47)(48)(49). Conversely, primed hPSCs displayed negligible expression of SUSD2 compared to naïve hPSCs (fig.…”

Section: Ila Naïve Hpsc Lines Are Derived From the Primed Source Line...supporting

confidence: 90%

Female naïve human pluripotent stem cells carry X chromosomes with Xa-like and Xi-like folding conformations

et al. 2023

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Three-dimensional (3D) genomics shows immense promise for studying X chromosome inactivation (XCI) by interrogating changes to the X chromosomes’ 3D states. Here, we sought to characterize the 3D state of the X chromosome in naïve and primed human pluripotent stem cells (hPSCs). Using chromatin tracing, we analyzed X chromosome folding conformations in these cells with megabase genomic resolution. X chromosomes in female naïve hPSCs exhibit folding conformations similar to the active X chromosome (Xa) and the inactive X chromosome (Xi) in somatic cells. However, naïve X chromosomes do not exhibit the chromatin compaction typically associated with these somatic X chromosome states. In H7 naïve human embryonic stem cells, XIST accumulation observed on damaged X chromosomes demonstrates the potential for naïve hPSCs to activate XCI-related mechanisms. Overall, our findings provide insight into the X chromosome status of naïve hPSCs with a single-chromosome resolution and are critical in understanding the unique epigenetic regulation in early embryonic cells.

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Section: Ila Naïve Hpsc Lines Are Derived From the Primed Source Line...supporting

confidence: 90%

Female naïve human pluripotent stem cells carry X chromosomes with Xa-like and Xi-like folding conformations

et al. 2023

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“…iPSC generation and characterization. Patient iPSC were generated and characterized as described (51). Reagents are listed in Supplemental Table 3.…”

Section: Patientsmentioning

confidence: 99%

Base editing repairs an SGCA mutation in human primary muscle stem cells

et al. 2021

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“…The iPSCs were generated and characterized as described previously [ 33 , 34 ] from peripheral blood mononuclear cells. For SID, cells were split with 0.5 mM EDTA (Thermo Fisher Sci., Waltham, MA, USA) to iPSC aggregates with 5–8 cells and seeded the cells in 6× well plate coated with hESC-grade Matrigel (Corning, New York, NY, USA) to obtain 30–40% confluency the next day.…”

Section: Methodsmentioning

confidence: 99%

LMNA Co-Regulated Gene Expression as a Suitable Readout after Precise Gene Correction

Wang

Krause

Escobar

et al. 2022

IJMS

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LMNA-related muscular dystrophy is an autosomal-dominant progressive disorder caused by mutations in LMNA. LMNA missense mutations are becoming correctable with CRISPR/Cas9-derived tools. Evaluating the functional recovery of LMNA after gene editing bears challenges as there is no reported direct loss of function of lamin A/C proteins in patient-derived cells. The proteins encoded by LMNA are lamins A/C, important ubiquitous nuclear envelope proteins but absent in pluripotent stem cells. We induced lamin A/C expression in induced pluripotent stem cells (iPSCs) of two patients with LMNA-related muscular dystrophy, NM_170707.4 (LMNA): c.1366A > G, p.(Asn456Asp) and c.1494G > T, p.(Trp498Cys), using a short three-day, serum-induced differentiation protocol and analyzed expression profiles of co-regulated genes, examples being COL1A2 and S100A6. We then performed precise gene editing of LMNA c.1366A > G using the near-PAMless (PAM: protospacer-adjacent motif) cytosine base editor. We show that the mutation can be repaired to 100% efficiency in individual iPSC clones. The fast differentiation protocol provided a functional readout and demonstrated increased lamin A/C expression as well as normalized expression of co-regulated genes. Collectively, our findings demonstrate the power of CRISPR/Cas9-mediated gene correction and effective outcome measures in a disease with, so far, little perspective on therapies.

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Generation of two human induced pluripotent stem cell lines derived from myoblasts (MDCi014-A) and from peripheral blood mononuclear cells (MDCi014-B) from the same donor

Cited by 7 publications

References 4 publications

Female naïve human pluripotent stem cells carry X chromosomes with Xa-like and Xi-like folding conformations

Female naïve human pluripotent stem cells carry X chromosomes with Xa-like and Xi-like folding conformations

Base editing repairs an SGCA mutation in human primary muscle stem cells

LMNA Co-Regulated Gene Expression as a Suitable Readout after Precise Gene Correction

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