Version 1
: Received: 13 June 2023 / Approved: 13 June 2023 / Online: 14 June 2023 (08:01:58 CEST)
Version 2
: Received: 16 September 2023 / Approved: 18 September 2023 / Online: 18 September 2023 (03:58:54 CEST)
Yeganeh, B.; Yeganeh, A.; Malone, K.; Beug, S.T.; Jankov, R.P. Suspension-Induced Stem Cell Transition: A Non-Transgenic Method to Generate Adult Stem Cells from Mouse and Human Somatic Cells. Cells2023, 12, 2508.
Yeganeh, B.; Yeganeh, A.; Malone, K.; Beug, S.T.; Jankov, R.P. Suspension-Induced Stem Cell Transition: A Non-Transgenic Method to Generate Adult Stem Cells from Mouse and Human Somatic Cells. Cells 2023, 12, 2508.
Yeganeh, B.; Yeganeh, A.; Malone, K.; Beug, S.T.; Jankov, R.P. Suspension-Induced Stem Cell Transition: A Non-Transgenic Method to Generate Adult Stem Cells from Mouse and Human Somatic Cells. Cells2023, 12, 2508.
Yeganeh, B.; Yeganeh, A.; Malone, K.; Beug, S.T.; Jankov, R.P. Suspension-Induced Stem Cell Transition: A Non-Transgenic Method to Generate Adult Stem Cells from Mouse and Human Somatic Cells. Cells 2023, 12, 2508.
Abstract
Adult stem cells (ASCs) can be cultured with difficulty from most tissues, often requiring chemical or transgenic modification to achieve adequate quantities. We show here that mouse primary fibroblasts grown in suspension change from the elongated and flattened morphology observed under standard adherent culture conditions generating rounded cells with large nuclei and scant cytoplasm expressing the mesenchymal stem cell (MSC) marker (Sca1; Ly6A) within 24hrs. Based on this initial observation, we describe here a suspension culture method that, irrespective of the lineage used, mouse fibroblast, primary human somatic cells (fibroblasts, hepatocytes and keratinocytes), is capable of generating a high yield of cells in spheroid form which display expression of ASCs surface markers, circumventing the anoikis which often occurs at this stage. Moreover, mouse fibroblasts-derived spheroids can be differentiated into adipogenic and osteogenic lineages. Analysis of single cell RNA sequence data identified 8 distinct cell clusters with one in particular comprising approximately 10% of the cells showing high levels of proliferative capacity expressing high levels of genes related to MSCs and self-renewal as well as extracellular matrix (ECM). We believe the rapid, high-yield generation of proliferative, multi-potent ASC-like cells by the process we term suspension-induced stem cell transition (SIST) could have significant implications for regenerative medicine.
Biology and Life Sciences, Cell and Developmental Biology
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