Effect of 17β-estradiol on the in vitro differentiation of murine embryonic stem cells into the osteogenic lineage

Abstract

Embryonic stem (ES) cells have the potential to differentiate into various cell types of the three germ layers. They are therefore a useful cell source for transplantation and tissue engineering. In the present paper, we studied the influences of ascorbic acid (AA), dexamethasone (Dex), and 17beta-estradiol (E(2)) on the osteogenic differentiation of ES cells. Differentiation into the osteoblastic phenotype was demonstrated by the appearance of osteoblastic markers such as alkaline phosphatase (ALP), the transcription factor core binding factor alpha 1 (Cbfa1), and osteocalcin, which were detected by immunohistochemistry. Bone nodule formation, including the deposition of collagen fibrils and matrix mineralization, was studied by transmission electron microscopy. In all our cultures, a progressive upregulation of ALP activity was observed, followed by a decline after 21 d of culture. Cbfa1 was first detected after 14 d in culture and increased during the culture time. The addition of E(2) resulted in a decrease in the formation of bone-like nodules in the embryoid bodies (EBs) compared with the EBs cultured in the presence of AA and AA supplemented with Dex. An increased osteocalcin concentration was observed in the EBs cultured with Dex and E(2) compared with the EBs cultured in a control medium. EBs cultured in the presence of E(2) resulted in a culture with a high amount of osteoblast-like cells not entrapped in bone-like nodules, creating the possibility to obtain a purified osteoblast population for bone tissue engineering.

Stimulatory effect of 17β-estradiol on osteogenic differentiation potential of rat adipose tissue-derived stem cells

“…Enhanced ALP expression in response to E 2 by osteoblasts and stem cells was demonstrated in some recent studies (Qu et al 1998;Holzer et al 2002;Tielens et al 2008;Hoemann et al 2009). Furthermore, E 2 probably acts not only to control the ALP activity of cells expressing the enzyme, but also the proliferation rate of ALP-expressing cells in the marrow stromal cell population (Holzer et al 2002).…”

“…Bodine et al (1998) reported increased levels of estrogen receptor during osteoblast differentiation in primary cultures of rat calvarial osteoblasts. More recently, it has been suggested that E 2 may also effectively enhance the differentiation capacity of both embryonic stem cells and BMSCs into ostoeblasts in vitro (Qu et al 1998;Hong et al 2006;Tielens et al 2008).…”

GABAAand GABABreceptors of distinct properties affect oppositely the proliferation of mouse embryonic stem cells through synergistic elevation of intracellular Ca2+

“…At least 3 wells were used per treatment per assay. AP histochemistry was performed as described previously (36).…”

Stimulating Effect of a Novel Synthesized Sulfonamido-Based Gallate ZXHA-TC on Primary Osteoblasts

“… 32 Commonly used as a marker of osteogenesis, ALP activity increases during the differentiation stage and declines when mineralization occurs. 38 , 39 In this study, ALP activity in the ZXHA-TC-treated groups increased early yet decreased later ( Fig. 7A ), contrary to the control, in which a continuous increase was observed during the whole period.…”