1996
DOI: 10.1002/(sici)1097-0185(199605)245:1<25::aid-ar5>3.3.co;2-j View full text |Buy / Rent full text
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Abstract: Backgmund: we reported that when Meckel's cartilage was transplanted ectopically, chondrocytes transformed into osteocyte-like cells accompanying the extracellular calcified matrix. However, we could not determine whether the osteocyte-like cells were derived from host tissues or from Meckel's cartilage itself. Therefore, we examined whether the Meckel's cartilage chondrocytes, which have a retrogressive ultimate fate, are capable of inducing the observed calcification and further transform into osteocyte-like… Show more

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“…Meckel's chondrocytes were isolated enzymatically from 17-day embryos of ddY mice, as described elsewhere (Ishizeki et al 1996b(Ishizeki et al , 1996c. The chondrocytes were suspended in alpha-modified Eagle's medium (α-MEM; Flow Laboratories, Irvine, Scotland; designated the control medium), supplemented with 10% fetal bovine serum (FBS; Mitsubishi Kasei, Tokyo, Japan), 0.03 mg/ml L-ascorbic acid phosphate magnesium salt n-hydrate ((C 6 H 6 O 9 P)·3/2Mg·nH 2 O (Wako Pure Chemical Industries, Osaka, Japan), 60 µg/ml kanamycin (Meiji Seika, Tokyo, Japan), and 3 mM β-glycerophosphate (Nacalai Tesque, Kyoto, Japan).…”
Section: Cell Culturementioning
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“…Meckel's chondrocytes were isolated enzymatically from 17-day embryos of ddY mice, as described elsewhere (Ishizeki et al 1996b(Ishizeki et al , 1996c. The chondrocytes were suspended in alpha-modified Eagle's medium (α-MEM; Flow Laboratories, Irvine, Scotland; designated the control medium), supplemented with 10% fetal bovine serum (FBS; Mitsubishi Kasei, Tokyo, Japan), 0.03 mg/ml L-ascorbic acid phosphate magnesium salt n-hydrate ((C 6 H 6 O 9 P)·3/2Mg·nH 2 O (Wako Pure Chemical Industries, Osaka, Japan), 60 µg/ml kanamycin (Meiji Seika, Tokyo, Japan), and 3 mM β-glycerophosphate (Nacalai Tesque, Kyoto, Japan).…”
Section: Cell Culturementioning
“…After being washed with PBS, samples were stained with 1% Alizarin red S (Kanto Chemical, Tokyo, Japan) or incubated for 20-30 min with a 5% solution of silver nitrate for von Kossa's staining, as described previously (Ishizeki et al 1996b(Ishizeki et al , 1999.…”
Section: Staining With Alizarin Red and Von Kossa's Stainmentioning
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“…On the other hand, Akisaka (1982) demonstrated thiamine pyrophosphatase activity, an osteoblastic marker, in the Golgi apparatus of hypertrophic chondrocytes in degenerating Meckel's cartilage, and proposed that these chondrocytes may survive and transdifferentiate to bone cells. Furthermore, Ishizeki et al (1996aIshizeki et al ( , 1996bIshizeki et al ( , 1996cIshizeki et al ( , 1998Ishizeki et al ( , 1999 demonstrated, using a cell culture model, that the chondrocytes of Meckel's cartilage can transdifferentiate in vitro to osteogenic cells as characterized by their electron microscopic morphology and production of type I collagen. Whatever the fate of the chondrocytes in Meckel's cartilage, the degenerating Meckel's cartilage differs from the cartilage undergoing endochondral ossification in that the extensive vascularization that introduces osteogenic cells into the cartilage matrix is lacking in the former (Ishizeki et al 1999).…”
Section: Discussionmentioning
“…Meckel's cartilage was dissected out on day 17 of gestation (day 0 = day of appearance of a vaginal plug) and was placed in phosphate-buffered saline (PBS) that contained 0.2% glucose (PBS-G). Chondrocytes were isolated with a mixture of enzymes, namely, 0.1% hyaluronidase (type I-S; Sigma Chemical Co., St. Louis, Mo., USA) and 0.15% collagenase (type II; Worthington Biochemical Corp., Freehold, N.J., USA), as described previously (Ishizeki et al 1996b(Ishizeki et al , 1998a.…”
Section: Preparation Of Cellsmentioning