Automated in-process characterization and selection of cell-clones for quality and efficient cell manufacturing


Delivery of safe, effective and reliable cellular therapies, whether based on mesenchymal stromal cells (MSCs) or induced pluripotent stem cells (iPSCs), demand standardization of cell culture protocols. There is a need to develop automation platform that enables the users to generate culture expanded human cell populations that improves the quality and reduces batch-to-batch variation with respect to biological potential. Cell X™ robot was designed to address these current challenges in the cell fabrication industry. It utilizes non-invasive large field of view quantitative image analysis to guide an automated process of targeted “biopsy” (cells or media), “picking” (selection) of desired cells or colonies, or “weeding” (removal) of undesired cells, thus providing an unprecedented ability to acquire quantitative measurement in a complex heterogeneous cell environment “in process” and then to act on those measurements to define highly reproducible methods for cell and colony “management” based on application specific critical quality attributes to improve the quality of the manufactured cell lines and cell products.

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  1. ASTM F2944-12 (2012) Standard test method for automated colony forming unit (CFU) assays—image acquisition and analysis method for enumerating and characterizing cells and colonies in culture.

  2. Cerbini T, Luo Y, Rao MS, Zou J (2015) Transfection, selection, and colony-picking of human induced pluripotent stem cells TALEN-targeted with a GFP gene into the AAVS1 safe harbor. J Vis Exp. 96:1–9.

    CAS  Article  Google Scholar 

  3. Chen KG, Mallon BS, McKay RDG, Robey PG (2014) Human pluripotent stem cell culture: considerations for maintenance, expansion, and therapeutics. Cell Stem Cell 14:13–26.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. Galipeau J, Sensébé L (2018) Mesenchymal stromal cells: clinical challenges and therapeutic opportunities. Cell Stem Cell 22:824–833.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  5. Haasters F, Prall WC, Anz D et al (2009) Morphological and immunocytochemical characteristics indicate the yield of early progenitors and represent a quality control for human mesenchymal stem cell culturing. J Anat 214:759–767.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Haupt S, Grützner J, Rath BH, Möhlig H, Brüstle O (2009) Automated selection and collection of pluripotent stem cell colonies using the Cell CelectorTM. Nat Methods 6:3–4.

    Article  Google Scholar 

  7. Kurtz A, Seltmann S, Bairoch A et al (2018) A standard nomenclature for referencing and authentication of pluripotent stem cells. Stem Cell Rep 10:300–313.

    CAS  Article  Google Scholar 

  8. Kutepatil O (2017) Precision medicine market to witness astonishing developments over 2017–2024, U. S. to transpire as a commercially viable ground for the industry growth. Fractovia

  9. Kwee E, Herderick EE, Adams T et al (2017) Integrated colony imaging, analysis, and selection device for regenerative medicine. SLAS Technol Transl Life Sci Innov. 22:217–223.

    Article  Google Scholar 

  10. Mantripragada VP, Bova W, Boehm C et al (2018a) Primary cells isolated from human knee cartilage reveal decreased prevalence of progenitor cells but comparable biological potential during osteoarthritic disease progression. J Bone Joint Surg Am 100:1771–1780

    CAS  Article  Google Scholar 

  11. Mantripragada VP, Bova WA, Boehm C et al (2018b) Progenitor cells from different zones of human cartilage and their correlation with histopathological osteoarthritis progression. J Orthop Res 36:1728–1738.

    CAS  Article  PubMed  Google Scholar 

  12. Mantripragada VP, Piuzzi NS, Bova WB, et al (2019) Donor-matched comparison of chondrogenic progenitors resident in human infrapatellar fat pad, synovium and periosteum and correlation to patient age and gender-implications for cartilage repair. Connect Tissue Res 60:597–610

    CAS  Article  Google Scholar 

  13. Mendicino M, Bailey AM, Wonnacott K, Puri RK, Bauer SR (2014) MSC-based product characterization for clinical trials: an FDA perspective. Cell Stem Cell 14:141–145.

    CAS  Article  PubMed  Google Scholar 

  14. Ng M, Song S, Piuzzi N et al (2017) Stem cell industry update: 2012 to 2016 reveals accelerated investment, but market capitalization and earnings lag stem cell industry update. Cytotherapy 19:1131–1139

    Article  Google Scholar 

  15. Piuzzi NS, Chahla J, Jiandong H et al (2017) Analysis of cell therapies used in clinical trials for the treatment of osteonecrosis of the femoral head: a systematic review of the literature. J Arthroplasty 32:2612–2618.

    Article  PubMed  Google Scholar 

  16. Piuzzi NS, Hussain ZB, Chahla J et al (2018) Variability in the preparation, reporting, and use of bone marrow aspirate concentrate in musculoskeletal disorders. J Bone Jt Surg 100:517–525.

    Article  Google Scholar 

  17. Powell KA, Nakamoto C, Villarruel S, Boehm C, Muschler G (2007) Quantitative image analysis of connective tissue progenitors. Anal Quant Cytol Histol. 29:112–121

    PubMed  Google Scholar 

  18. Powell K, Kwee E, Nutter B et al (2016) Variability in subjective review of umbilical cord blood colony forming unit assay. Cytom Part B Clin Cytom. 90:517–524.

    CAS  Article  Google Scholar 

  19. Qadan MA, Piuzzi NS, Boehm C et al (2018) Variation in primary and culture-expanded cells derived from connective tissue progenitors in human bone marrow space, bone trabecular surface and adipose tissue. Cytotherapy. 20:343–360.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  20. Ramkumar PN, Navarro SM, Haeberle HS et al (2017) Cellular therapy injections in today’s orthopedic market: a social media analysis. Cytotherapy 19:1392–1399

    Article  Google Scholar 

  21. Russell KC, Phinney DG, Lacey MR, Barrilleaux BL, Meyertholen KE, O Connor KC (2010) In vitro high-capacity assay to quantify the clonal heterogeneity in trilineage potential of mesenchymal stem cells reveals a complex hierarchy of lineage commitment. Stem Cells. 28:788–798.

    CAS  Article  PubMed  Google Scholar 

  22. Schwarz BA, Cetinbas M, Clement K et al (2018) Prospective isolation of poised iPSC intermediates reveals principles of cellular reprogramming. Cell Stem Cell 23:289–305.e5.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  23. Seltmann S, Lekschas F, Müller R et al (2016) hPSCreg-the human pluripotent stem cell registry. Nucleic Acids Res 44:D757–D763.

    CAS  Article  PubMed  Google Scholar 

  24. Stoltz J-F, de Isla N, Li YP et al (2015) Stem cells and regenerative medicine: myth or reality of the 21th century. Stem Cells Int 2015:734731.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  25. Takahashi K, Tanabe K, Ohnuki M et al (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131:861–872.

    CAS  Article  PubMed  Google Scholar 

  26. Vitale AM, Matigian NA, Ravishankar S et al (2012) Variability in the generation of induced pluripotent stem cells: importance for disease modeling. Stem Cells Transl Med. 1:641–650.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  27. Wrighton KH (2017) Stem cells: the different flavours of iPS cells. Nat Rev Genet 18:394.

    CAS  Article  PubMed  Google Scholar 

  28. Zoldan K, Arnold A, Stolzing A (2010) Automated harvest of induced pluripotent stem cell Ccolonies and colony fractions using the cell separation robot cell celector. Advert Feature Nat Methods

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Cleveland Clinic and Parker Hannifin have collaborated to create Cell X™ Platform. Biospherix (Lacona, New York) provided active support enabling the design of the Cell X™ Work Space, based on the CytoCentric™ principles. iPS-derived cardiomyocytes were studied in collaboration with David Van-Wagoner.

The work was supported by funding from NIH (R21AR067357), Lisa Dean Moseley Foundation and NIH-NCAI (NCAI-17-7-APP-CCF-Muschler) on which GFM is the Principal Investigator.

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Correspondence to Venkata P. Mantripragada.

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GFM and KP are named inventors on two issued and one pending patent that have been licensed by Cell X Technologies.

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Mantripragada, V.P., Luangphakdy, V., Hittle, B. et al. Automated in-process characterization and selection of cell-clones for quality and efficient cell manufacturing. Cytotechnology (2020).

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  • Automation
  • Stem-cell manufacturing
  • Cell-picking
  • Imaging
  • Performance-based cell selection
  • Stem cells