title: Isolation of muscle-derived stem/progenitor cells based on adhesion characteristics to collagen-coated surfaces.
abstract: Our lab developed and optimized a method, known as the modified pre-plate technique, to isolate stem/progenitor cells from skeletal muscle. This method separates different populations of myogenic cells based on their propensity to adhere to a collagen I-coated surface. Based on their surface markers and stem-like properties, including self-renewal, multi-lineage differentiation, and ability to promote tissue regeneration, the last cell fraction or slowest to adhere to the collagen-coated surface (pre-plate 6; pp6) appears to be early, quiescent progenitor cells termed muscle-derived stem/progenitor cells (MDSPCs). The cell fractions preceding pp6 (pp1-5) are likely populations of more committed (differentiated) cells, including fibroblast- and myoblast-like cells. This technique may be used to isolate MDSPCs from skeletal muscle of humans or mice regardless of age, sex or disease state, although the yield of MDSPCs varies with age and health. MDSPCs can be used for regeneration of a variety of tissues including bone, articular cartilage, skeletal and cardiac muscle, and nerve. MDSPCs are currently being tested in clinical trials for treatment of urinary incontinence and myocardial infarction. MDSPCs from young mice have also been demonstrated to extend life span and healthspan in mouse models of accelerated aging through an apparent paracrine/endocrine mechanism. Here we detail methods for isolation and characterization of MDSPCs.
authors: Department of Orthopaedic Surgery, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
authors: Mitra
authors: M
authors: Lavasani
authors: Aiping
authors: A
authors: Lu
authors: Seth D
authors: SD
authors: Thompson
authors: Paul D
authors: PD
authors: Robbins
authors: Johnny
authors: J
authors: Huard
authors: Laura J
authors: LJ
authors: Niedernhofer
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism*; D051379:Mice; D018482:Muscle, Skeletal / Q000166:cytology* / Q000502:physiology; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology
chemical_list: D003094:Collagen
references: Nat Commun. 2012 Jan 03;3:608
references: 22215083
references: Medicina (Kaunas). 2011;47(9):469-79
references: 22156603
references: Biochem Biophys Res Commun. 2004 May 7;317(3):893-901
references: 15081424
references: Stem Cells. 2003;21(5):536-45
references: 12968108
references: Cell. 2004 Nov 12;119(4):543-54
references: 15537543
references: Neurosurg Clin N Am. 2007 Jan;18(1):81-92, ix
references: 17244556
references: J Bone Joint Surg Am. 2001 Jul;83(7):1032-9
references: 11451972
references: Proc Natl Acad Sci U S A. 1999 Dec 7;96(25):14482-6
references: 10588731
references: Am J Physiol Renal Physiol. 2004 Oct;287(4):F621-7
references: 15198930
references: Biochem J. 2007 Jun 1;404(2):169-78
references: 17488235
references: Tissue Eng. 2001 Aug;7(4):395-404
references: 11506729
references: J Bone Joint Surg Am. 2004 Jan;86(1):80-91
references: 14711949
references: Eur J Biochem. 1988 May 16;174(1):103-10
references: 3371354
references: Mol Ther. 2002 Aug;6(2):169-78
references: 12161183
references: Urology. 2003 Nov;62(5):958-63
references: 14624934
references: Exp Cell Res. 2008 Apr 1;314(6):1266-80
references: 18282570
references: Pol Arch Med Wewn. 2010 Jul;120(7-8):282-9
references: 20693959
references: PLoS Biol. 2005 Apr;3(4):e87
references: 15757365
references: Nat Cell Biol. 2001 Sep;3(9):778-84
references: 11533656
references: Proc Natl Acad Sci U S A. 2008 Jan 29;105(4):1226-31
references: 18212116
references: Differentiation. 1996 Mar;60(1):47-57
references: 8935928
references: J Anat. 2005 Sep;207(3):197-208
references: 16185244
references: J Cell Biol. 2000 Sep 4;150(5):1085-100
references: 10973997
references: J Bone Miner Res. 2005 Nov;20(11):2017-27
references: 16234975
references: Trends Cardiovasc Med. 2002 Apr;12(3):115-20
references: 12007736
references: Hum Gene Ther. 2003 Nov 1;14(16):1535-46
references: 14577915
references: J Neurosci Res. 2002 Sep 15;69(6):894-907
references: 12205682
references: N Engl J Med. 2011 Nov 17;365(20):1853-6
references: 22087677
references: Gene Ther. 2005 Aug;12(16):1264-74
references: 15843810
references: Arthritis Rheum. 2006 Feb;54(2):433-42
references: 16447218
references: J Cell Biol. 2002 May 27;157(5):851-64
references: 12021255
references: Int Urogynecol J Pelvic Floor Dysfunct. 2008 Jun;19(6):881-3
references: 18204978
references: Arthritis Rheum. 2009 May;60(5):1390-405
references: 19404941
references: Nature. 2007 Oct 25;449(7165):1003-7
references: 17934449
references: Mol Ther. 2012 Jan;20(1):138-45
references: 22068427
references: Curr Opin Mol Ther. 2010 Aug;12(4):442-9
references: 20677095
references: Tissue Eng Part A. 2012 Feb;18(3-4):232-41
references: 21854253
references: Hum Gene Ther. 2001 Apr 10;12(6):619-28
references: 11426462
references: Mol Ther. 2009 Oct;17(10):1788-98
references: 19603004
references: Obes Res. 2003 Jan;11(1):5-16
references: 12529479
references: Int Urogynecol J Pelvic Floor Dysfunct. 2003 Feb;14(1):31-7; discussion 37
references: 12601514
references: Cell Prolif. 1995 Mar;28(3):137-44
references: 7734623
references: Mol Ther. 2005 Dec;12(6):1130-41
references: 16125468
references: J Orthop Res. 2007 Nov;25(11):1534-40
references: 17593537
references: Stem Cell Rev. 2006;2(3):203-12
references: 17625256
references: Urology. 2006 Aug;68(2):449-54
references: 16904482
references: Nat Cell Biol. 2003 Jul;5(7):640-6
references: 12792651
references: J Cell Sci Suppl. 1988;10:63-76
references: 3077943
references: J Bone Joint Surg Br. 2002 Jan;84(1):120-7
references: 11837817
references: J Clin Invest. 2002 Sep;110(6):751-9
references: 12235106
references: Cell Prolif. 2011 Feb;44(1):86-98
references: 21199013
references: Hum Gene Ther. 2006 Feb;17(2):180-92
references: 16454652
references: Science. 1987 Oct 16;238(4825):347-50
references: 3659917
references: Reprod Fertil Dev. 2001;13(1):23-9
references: 11545161
references: J Cell Sci. 2006 Jun 1;119(Pt 11):2204-13
references: 16684817
references: J Cell Biol. 2002 May 13;157(4):571-7
references: 11994315
references: Mol Biol Cell. 2005 Jul;16(7):3323-33
references: 15872085
references: J Cell Biol. 2001 Jan 22;152(2):335-48
references: 11266450
references: Urology. 2004 Apr;63(4):780-5
references: 15072911
references: Trends Mol Med. 2001 Jul;7(7):301-6
references: 11425638
references: Nature. 1999 Sep 23;401(6751):390-4
references: 10517639