Proliferation of germ cells and somatic cells in first trimester human embryonic gonads as indicated by S and S+G2+M phase fractions
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Proliferation of germ cells and somatic cells in first trimester human embryonic gonads as indicated by S and S+G2+M phase fractions. / Sørensen, Kirstina Pilekær; Lutterodt, M C; Mamsen, L S; Byskov, A G; Larsen, J K.
I: Cell Proliferation, Bind 44, Nr. 3, 06.2011, s. 224-33.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Proliferation of germ cells and somatic cells in first trimester human embryonic gonads as indicated by S and S+G2+M phase fractions
AU - Sørensen, Kirstina Pilekær
AU - Lutterodt, M C
AU - Mamsen, L S
AU - Byskov, A G
AU - Larsen, J K
N1 - © 2011 Blackwell Publishing Ltd.
PY - 2011/6
Y1 - 2011/6
N2 - OBJECTIVES: The number of germ cells and somatic cells in human embryonic and foetal gonads has previously been estimated by stereological methods, which are time- and labour-consuming with little information concerning cell proliferation. Here, we studied whether flow cytometry could be applied as an easier method, also enabling estimation of the fraction of cells in S or S+G(2)+M (SG(2) M) cell-cycle phases as indicators of cell proliferation.METHODS: Cell suspensions from 35 human embryonic gonads at days 37 to 68 post-conception (pc) were immunomagnetically sorted into C-KIT positive (germ) cells and negative (somatic) cells. They were stained for DNA content and analysed by flow cytometry. S and SG(2) M fractions could be measured for 13 of the female and 20 of the male gonads. The number of cells was estimated using fluorescent reference beads.RESULTS: During the period from 37 to 68 days pc, female germ and somatic cells had a stable S and SG(2) M fractions indicating steady growth of both subpopulations, whereas they decreased in both male germ and somatic cells. The number of germ and somatic cells estimated by flow cytometry was significantly lower than in stereological estimates, suggesting loss of cells during preparation.CONCLUSIONS: Cell proliferation as indicated by S and SG(2) M fractions could be estimated specifically for primordial germ and somatic cells. Estimation of total number of germ and somatic cells was not feasible.
AB - OBJECTIVES: The number of germ cells and somatic cells in human embryonic and foetal gonads has previously been estimated by stereological methods, which are time- and labour-consuming with little information concerning cell proliferation. Here, we studied whether flow cytometry could be applied as an easier method, also enabling estimation of the fraction of cells in S or S+G(2)+M (SG(2) M) cell-cycle phases as indicators of cell proliferation.METHODS: Cell suspensions from 35 human embryonic gonads at days 37 to 68 post-conception (pc) were immunomagnetically sorted into C-KIT positive (germ) cells and negative (somatic) cells. They were stained for DNA content and analysed by flow cytometry. S and SG(2) M fractions could be measured for 13 of the female and 20 of the male gonads. The number of cells was estimated using fluorescent reference beads.RESULTS: During the period from 37 to 68 days pc, female germ and somatic cells had a stable S and SG(2) M fractions indicating steady growth of both subpopulations, whereas they decreased in both male germ and somatic cells. The number of germ and somatic cells estimated by flow cytometry was significantly lower than in stereological estimates, suggesting loss of cells during preparation.CONCLUSIONS: Cell proliferation as indicated by S and SG(2) M fractions could be estimated specifically for primordial germ and somatic cells. Estimation of total number of germ and somatic cells was not feasible.
KW - Alkaline Phosphatase/metabolism
KW - Cell Proliferation
KW - Cell Separation
KW - Female
KW - Flow Cytometry/methods
KW - Fluorescent Dyes/pharmacology
KW - Germ Cells/cytology
KW - Gonads/cytology
KW - Humans
KW - Immunomagnetic Separation
KW - Male
KW - Pregnancy
KW - Pregnancy Trimester, First
KW - Proto-Oncogene Proteins c-kit/biosynthesis
KW - U937 Cells
U2 - 10.1111/j.1365-2184.2011.00748.x
DO - 10.1111/j.1365-2184.2011.00748.x
M3 - Journal article
C2 - 21535263
VL - 44
SP - 224
EP - 233
JO - Cell Proliferation
JF - Cell Proliferation
SN - 0960-7722
IS - 3
ER -
ID: 203597567