scholarly journals Measurement of apoptosis in long-term cultures of human ovarian tissue

Reproduction ◽  
2001 ◽  
pp. 657-663 ◽  
Author(s):  
G Rahimi ◽  
E Isachenko ◽  
H Sauer ◽  
M Wartenberg ◽  
V Isachenko ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Culture Conditions ◽  
Apoptotic Cells ◽  
Experimental Conditions ◽  
Tissue Samples ◽  
Long Term Culture ◽  
Hormone Synthesis ◽  
Human Ovarian Tissue ◽  
Control Samples

At present, the long-term culture of ovarian tissue is problematic. The aim of this study was to measure apoptosis in long-term cultures of human ovarian tissue. Biopsies of human ovaries were cultured for 6 weeks. Samples were taken weekly for histological investigation. The apoptotic cells were marked with anti-caspase 3. Simultaneous to this experiment, other tissue samples were preincubated for 3 h with 1 micromol staurosporine l(-1), an inducer of apoptosis, and apoptosis was compared among samples. Furthermore, the proportion of lethal cells was determined weekly. After 6 weeks, 99% of the tissue samples showed an intact structure. They expanded in all directions on the floor of the multi-wells to form a monolayer. Apoptotic cells could be marked only sporadically (16.3 +/- 5.9 fluorescence (counts per 3600 microm(2))) after 6 weeks. After preincubation with staurosporine after the same period of culture, the proportion of apoptotic cells was significantly increased compared with that in untreated control samples (66.8 +/- 14.5 versus 16.3 +/- 5.9%, respectively; P < 0.05). Under the same experimental conditions, the proportion of lethal cells was 3.6 +/- 0.9, 3.9 +/- 2.1 and 5.2 +/- 1.5% for weeks 1, 3 and 6, respectively. After preincubation with 1 micromol staurosporine l(-1), the proportion of pyknotic cells after 6 weeks of culture was significantly higher (37.2 +/- 4.4%) than that in control samples (3.95 +/- 2.05%; P < 0.05). No significant increase in apoptosis in cultured human ovarian tissue after 6 weeks was observed compared with control tissues on day 1. These results indicate that under optimal culture conditions it is possible to cultivate human ovarian tissue long term. The influence of long-term culture on hormone synthesis and follicle maturity will be investigated further.

scholarly journals Impact of freezing and thawing of human ovarian tissue on follicular growth after long-term xenotransplantation

2011 ◽  
Vol 28 (12) ◽  
pp. 1157-1165 ◽  
Author(s):  
Christiani A. Amorim ◽  
Anu David ◽  
Marie-Madeleine Dolmans ◽  
Alessandra Camboni ◽  
Jacques Donnez ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Follicular Growth ◽  
Freezing And Thawing ◽  
Human Ovarian Tissue

Slush nitrogen vitrification of human ovarian tissue does not alter gene expression and improves follicle health and progression in long-term in vitro culture

2018 ◽  
Vol 110 (7) ◽  
pp. 1356-1366 ◽  
Author(s):  
Vincenza Barbato ◽  
Roberto Gualtieri ◽  
Teresa Capriglione ◽  
Maria Michela Pallotta ◽  
Sabrina Braun ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Vitro Culture ◽  
Ovarian Tissue ◽  
Human Ovarian Tissue

HUMAN ENDOTHELIAL CELLS MODIFICATIONS DURING LONG TERM CULTURE

1987 ◽  
Author(s):  
M P Wautier ◽  
J L Wautier
Keyword(s):  
Endothelial Cells ◽  
Doubling Time ◽  
Umbilical Vein ◽  
Light And Electron Microscopy ◽  
Major Change ◽  
Culture Conditions ◽  
Human Endothelial Cells ◽  
Long Term Culture

The culture of human endothelial cells is largely used for vascular research. The possibility of developping long term culture of human endothelial cells (EC) raised the question regarding the identity after several passages. To further investigate this aspect we have cultured human umbilical vein EC until the 12th passage on fibronectin coated dishes supplemented with ECGF. We have studied the EC morphology by light and electron microscopy, the reactivity with 51Cr labelled platelets, and prostacyclin synthesis. Until the 6th passage no major change could be noted, except the occurence of rare large EC and a reduction in the doubling time between 2nd and 5th passage. After the 7th passage up to the 10th EC became more elongated and did not grow in strict monolayer. The number of vacuoles and mitochondria increased as well as the doubling time. After the 12th passage the EC were still viable but proliferated very slowly. The adhesion of radiolabelled platelets dramatically increased (150%) and PGI2 production significantly decreased (6 Keto PGF1α : 1st passage 13±2.5 ng; 6th passage 0.33±0.27 ng/106 EC). In our culture conditions EC kept most of their original characteristics up to the 6th passage but then lost some of them. At any passage EC contained Weibel Palade bodies and von Willebrand factor. We can conclude that after the 7th passage EC in culture are different from the original cells and could possibly represent an in vitro model of EC ageing.

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