Proliferation of germ cells and somatic cells in first trimester human embryonic gonads as indicated by S and S+G2+M phase fractions

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

SN - 0960-7722

VL - 44

SP - 224

EP - 233

JO - Cell Proliferation

JF - Cell Proliferation

IS - 3

ER -