Archives

  • 2018-07
  • 2018-10
  • 2018-11
  • 2019-04
  • 2019-05
  • 2019-06
  • 2019-07
  • 2019-08
  • 2019-09
  • 2019-10
  • 2019-11
  • 2019-12
  • 2020-01
  • 2020-02
  • 2020-03
  • 2020-04
  • 2020-05
  • 2020-06
  • 2020-07
  • 2020-08
  • 2020-09
  • 2020-10
  • 2020-11
  • 2020-12
  • 2021-01
  • 2021-02
  • 2021-03
  • 2021-04
  • 2021-05
  • 2021-06
  • 2021-07
  • 2021-08
  • 2021-09
  • 2021-10
  • 2021-11
  • 2021-12
  • 2022-01
  • 2022-02
  • 2022-03
  • 2022-04
  • 2022-05
  • 2022-06
  • 2022-07
  • 2022-08
  • 2022-09
  • 2022-10
  • 2022-11
  • 2022-12
  • 2023-01
  • br Materials and methods br Author disclosure statement br A

    2018-10-20


    Materials and methods
    Author disclosure statement
    Acknowledgments This work was supported by the UK Medical Research Council grants G0701172 and G0801061. We thank Dr. Yacoub Khalaf, Director of the Assisted Conception Unit of Guy\'s and St Thomas\' NHS Foundation Trust and his staff for supporting the research program. We are especially indebted to Prof Peter Braude and patients who donated embryos.
    Resource table
    Resource details
    We generated KCL029 clinical grade hESC line following protocols, established previously (Ilic et al., 2012; Stephenson et al., 2012). The expression of the pluripotency markers was tested after freeze/thaw cycle (Fig. 2). Differentiation potential into three germ layers was verified in vitro (Fig. 3).
    Materials and methods
    Author disclosure statement
    Acknowledgments This work was supported by the UK Medical Research Council grants G0701172 and G0801061. We thank Dr. Yacoub Khalaf, Director of the Assisted Conception Unit of Guy\'s and St Thomas\' NHS Foundation Trust and his staff for supporting the research program. We are especially indebted to Prof Peter Braude and to the patients who donated embryos.
    Resource table
    Resource details BJNhem20–OCIAD1-OV line was generated by transfection of BJNhem20 human embryonic stem cells (hESCs) with pCAG-OCIAD1 construct using microporation technique with the conditions of 1100V, 20ms pulse width and 2pulses. The pCAG-OCIAD1 construct was generated by cloning OCIAD1 Open Reading Frame amplicon from molar to millimolar 1–245 between Xho1 and Not1 sites using appropriate primers. A stable hESC line was generated after subjecting these transfected cells to puromycin selection for two weeks.
    Materials and methods
    Verification of karyotype
    Acknowledgments
    This project was funded by the Department of Biotechnology, Government of India and the Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore.
    Resource table
    Resource details
    We generated KCL031 clinical grade hESC line following protocols, established previously (Ilic et al., 2012; Stephenson et al., 2012), and now adapted to cGMP conditions. The expression of the pluripotency markers was tested after freeze/thaw cycle (Fig. 1). Differentiation potential into three germ layers was verified in vitro (Fig. 2) and in vivo (Fig. 3), as well as targeted differentiation into cardiac myocytes (Fig. 4). Molecular karyotyping using array comparative genomic hybridization aCGH identified deletion at 7q22.3 (105,465,968–105,516,305). Whole-genome single nucleotide polymorphism (SNP) array analysis detected loss at 8q24.23 (136,718,037–136,837,768) (Canham et al., 2015). The gain contains no genes and it has been also reported previously to occur in healthy individuals from worldwide population (Macdonald et al., 2014). Estimated frequency in the human population is 3.85% (Canham et al., 2015).
    Materials and methods
    Author disclosure statement
    Acknowledgments This work was supported by the UK Medical Research Council grants G0701172 and G0801061. We thank Dr. Yacoub Khalaf, Director of the Assisted Conception Unit of Guy\'s and St Thomas\' NHS Foundation Trust and his staff for supporting the research program. We are especially indebted to Prof Peter Braude and patients who donated embryos.
    Resource Table
    Resource Details Human skin fibroblasts, obtained by skin biopsy of a symptomatic, male 52-year-old spinocerebellar type 2 (SCA2) patient (anonymized as H196), were reprogrammed using episomal vectors carrying transcripts for human OCT4, SOX2, KLF4, L-MYC, LIN28, and small hairpin RNA for TP53 (Okita et al., 2011). The clone described in this publication was termed H196 clone (c) 7. Absence of the reprogramming plasmids was confirmed by quantitative PCR (qPCR) on genomic DNA (Fig. 1A).