title: Mitotic Diversity in Homeostatic Human Interfollicular Epidermis.
abstract: Despite decades of skin research, regulation of proliferation and homeostasis in human epidermis is still insufficiently understood. To address the role of mitoses in tissue regulation, we utilized human long-term skin equivalents and systematically assessed mitoses during early epidermal development and long-term epidermal regeneration. We now demonstrate four different orientations: (1) horizontal, i.e., parallel to the basement membrane (BM) and suggestive of symmetric divisions; (2) oblique with an angle of 45°-70°; or (3) perpendicular, suggestive of asymmetric division. In addition, we demonstrate a fourth substantial fraction of suprabasal mitoses, many of which are committed to differentiation (Keratin K10-positive). As verified also for normal human skin, this spatial mitotic organization is part of the regulatory program of human epidermal tissue homeostasis. As a potential marker for asymmetric division, we investigated for Numb and found that it was evenly spread in almost all undifferentiated keratinocytes, but indeed asymmetrically distributed in some mitoses and particularly frequent under differentiation-repressing low-calcium conditions. Numb deletion (stable knockdown by CRISPR/Cas9), however, did not affect proliferation, neither in a three-day follow up study by life cell imaging nor during a 14-day culture period, suggesting that Numb is not essential for the general control of keratinocyte division.
authors.affiliation: Department of Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany. katharinanoeske@gmx.de.
authors.forename: Katharina
authors.initials: K
authors.lastname: Nöske
authors.affiliation: Department of Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany. hj.stark@dkfz.de.
authors.forename: Hans-Jürgen
authors.initials: HJ
authors.lastname: Stark
authors.affiliation: Department of Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany. L.Nevaril@dkfz.de.
authors.forename: Leonard
authors.initials: L
authors.lastname: Nevaril
authors.affiliation: Department of Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany. manuel.berning@googlemail.com.
authors.forename: Manuel
authors.initials: M
authors.lastname: Berning
authors.affiliation: Department of Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany. L.Langbein@dkfz.de.
authors.forename: Lutz
authors.initials: L
authors.lastname: Langbein
authors.affiliation: Department of RNA Biology and Cancer, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany. A.Goyal@dkfz.de.|Institute of Pathology, University Hospital Heidelberg, Im Neuenheimer Feld 224, Heidelberg 69120, Germany. A.Goyal@dkfz.de.
authors.forename: Ashish
authors.initials: A
authors.lastname: Goyal
authors.affiliation: Department of RNA Biology and Cancer, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany. S.Diederichs@dkfz.de.|Division of Cancer Research, Department of Thoracic Surgery, Medical Center, University of Freiburg-Faculty of Medicine, University of Freiburg, Breisacher Str. 115, Freiburg 79106, Germany. S.Diederichs@dkfz.de.|German Cancer Consortium (DKTK), Freiburg 79106, Germany. S.Diederichs@dkfz.de.|Institute of Pathology, University Hospital Heidelberg, Im Neuenheimer Feld 224, Heidelberg 69120, Germany. S.Diederichs@dkfz.de.
authors.forename: Sven
authors.initials: S
authors.lastname: Diederichs
authors.affiliation: Department of Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany. p.boukamp@dkfz.de.|IUF-Leibniz Research Institute for Environmental Medicine, Düsseldorf 40225, Germany. p.boukamp@dkfz.de.
authors.forename: Petra
authors.initials: P
authors.lastname: Boukamp
mesh_terms: D060049:Asymmetric Cell Division; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D057809:HEK293 Cells; D006706:Homeostasis; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008938:Mitosis; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C113026:NUMB protein, human; D002118:Calcium
keywords: Numb; asymmetric mitosis; epidermal differentiation; human epidermis; long-term organotypic culture model; suprabasal mitosis
references.citation: J Cell Biol. 2000 May 15;149(4):851-62
references.pmid: 10811826
references.citation: Cell Transplant. 2016;25(2):353-64
references.pmid: 25994834
references.citation: Curr Biol. 2004 Feb 3;14(3):R129-38
references.pmid: 14986688
references.citation: Curr Opin Cell Biol. 2004 Apr;16(2):195-205
references.pmid: 15196564
references.citation: Cell. 2004 Sep 3;118(5):635-48
references.pmid: 15339667
references.citation: J Cell Biol. 2004 Oct 25;167(2):215-21
references.pmid: 15492044
references.citation: Br J Dermatol. 1970 Mar;82(3):276-82
references.pmid: 5441760
references.citation: Biochem Biophys Res Commun. 1973 Feb 5;50(3):744-50
references.pmid: 4734678
references.citation: J Biol Chem. 1982 Jun 10;257(11):6064-72
references.pmid: 6281254
references.citation: Br J Dermatol. 1985 Jul;113(1):53-64
references.pmid: 2410005
references.citation: Cell. 1989 Jul 28;58(2):349-60
references.pmid: 2752427
references.citation: Cell. 1994 Feb 11;76(3):477-91
references.pmid: 8313469
references.citation: Neuron. 1996 Jul;17(1):43-53
references.pmid: 8755477
references.citation: Chromosoma. 1997 Nov;106(6):348-60
references.pmid: 9362543
references.citation: Cell. 2004 Nov 12;119(4):503-16
references.pmid: 15537540
references.citation: Am J Pathol. 2005 Jul;167(1):59-69
references.pmid: 15972952
references.citation: Nature. 2005 Sep 8;437(7056):275-80
references.pmid: 16094321
references.citation: Neuron. 2005 Nov 23;48(4):539-45
references.pmid: 16301171
references.citation: Dev Cell. 2005 Dec;9(6):855-61
references.pmid: 16326396
references.citation: Nat Med. 2005 Dec;11(12):1351-4
references.pmid: 16288281
references.citation: Biochim Biophys Acta. 2006 Aug;1758(8):1034-42
references.pmid: 16872579
references.citation: J Investig Dermatol Symp Proc. 2006 Sep;11(1):93-105
references.pmid: 17069016
references.citation: Nature. 2007 Mar 8;446(7132):185-9
references.pmid: 17330052
references.citation: Eur J Cell Biol. 2007 Dec;86(11-12):731-46
references.pmid: 17292509
references.citation: Nature. 2008 Jan 3;451(7174):76-80
references.pmid: 18172499
references.citation: Cell Stem Cell. 2007 Oct 11;1(4):371-81
references.pmid: 18371376
references.citation: PLoS One. 2008;3(10):e3338
references.pmid: 18833336
references.citation: Stem Cells. 2008 Oct;26(10):2506-15
references.pmid: 18653773
references.citation: Cell Stem Cell. 2009 May 8;4(5):427-39
references.pmid: 19427292
references.citation: Genes Dev. 2010 Oct 15;24(20):2239-63
references.pmid: 20952534
references.citation: J Cell Biol. 2010 Nov 29;191(5):915-22
references.pmid: 21098114
references.citation: Biochim Biophys Acta. 2011 Jan;1815(1):26-43
references.pmid: 20940030
references.citation: Nature. 2011 Feb 17;470(7334):353-8
references.pmid: 21331036
references.citation: Cell. 2011 Jul 22;146(2):334-334.e2
references.pmid: 21784251
references.citation: Int J Biol Sci. 2012;8(2):228-35
references.pmid: 22253566
references.citation: Int Rev Cell Mol Biol. 2012;295:199-232
references.pmid: 22449491
references.citation: Cancer Res. 2012 Aug 1;72(15):3864-72
references.pmid: 22593191
references.citation: Cell Calcium. 2013 Feb;53(2):102-11
references.pmid: 23140583
references.citation: EMBO Rep. 2013 Aug;14(8):704-10
references.pmid: 23797874
references.citation: Nat Biotechnol. 2013 Sep;31(9):827-32
references.pmid: 23873081
references.citation: J Cell Biol. 2013 Sep 16;202(6):887-900
references.pmid: 24019538
references.citation: Philos Trans R Soc Lond B Biol Sci. 2013;368(1629):20130016
references.pmid: 24062586
references.citation: Nat Protoc. 2013 Nov;8(11):2281-308
references.pmid: 24157548
references.citation: Dev Cell. 2013 Nov 11;27(3):319-30
references.pmid: 24183650
references.citation: Curr Opin Cell Biol. 2013 Dec;25(6):749-58
references.pmid: 24021274
references.citation: Sci Signal. 2014 Oct 21;7(348):ra101
references.pmid: 25336614
references.citation: Tissue Eng Part C Methods. 2015 Sep;21(9):958-70
references.pmid: 25837604
references.citation: J Cell Biol. 2015 Nov 23;211(4):845-62
references.pmid: 26598619
references.citation: Nat Rev Genet. 2002 Mar;3(3):199-209
references.pmid: 11972157