TISSUE CULTURE OF EQUINE OVARIAN CELL TYPES: CULTURE METHODS AND MORPHOLOGY

1969 ◽  
Vol 43 (3) ◽  
pp. 381-NP ◽  
Author(s):  
CORNELIA P. CHANNING
Keyword(s):  
Tissue Culture ◽  
Cell Division ◽  
Granulosa Cells ◽  
Lipid Droplets ◽  
Horse Serum ◽  
Cell Types ◽  
Culture Methods ◽  
Ovarian Cell ◽  
Chromatin Material ◽  
Fibroblastic Cells

SUMMARY Equine granulosa and other cell types have been cultured for up to 70 days in a medium containing 15% horse serum, 30% medium '199' and 55% Hanks's solution. Granulosa cells harvested from large (3–6 cm. in diameter) vascular follicles of mares in oestrus grew in epithelioid colonies. Cell division and hypertrophy lasted for 7 days with the greatest amount of hypertrophy occurring during the first 3–4 days in culture. Within 3 days the cytoplasm increased in size and acquired eosinophilic granules and lipid droplets; nuclei acquired distinct chromatin material and nucleoli. Cells harvested from small follicles grew in fibroblastic stellate colonies and underwent hypertrophy and hyperplasia but did not show any of the cytoplasmic characteristics of the cultures of granulosa cells obtained from the large follicles. Heterogeneous fibroblastic cells grew from explants of stromal and thecal tissue from days 5 to 20 of culture.

STUDIES ON TISSUE CULTURE OF EQUINE OVARIAN CELL TYPES: EFFECT OF GONADOTROPHINS AND STAGE OF CYCLE ON STEROIDOGENESIS

1969 ◽  
Vol 43 (3) ◽  
pp. 415-425 ◽  
Author(s):  
CORNELIA P. CHANNING
Keyword(s):  
Tissue Culture ◽  
Cell Cultures ◽  
Granulosa Cells ◽  
Horse Serum ◽  
Cell Types ◽  
Phase Cell ◽  
Medium Change ◽  
Midluteal Phase

SUMMARY Granulosa cells were harvested from follicles of mares at various stages of the oestrous cycle and maintained in a tissue culture medium containing 15% horse serum, 30% medium '199' and 55% Hanks's solution. Between days 4 and 10 of culture the granulosa cells harvested from small follicles (1–2 cm. diam.) of mares in the midluteal phase of the cycle secreted an average of 0·36 pg. progesterone/cell/day. Cells harvested from large follicles of mares in the late and/or early oestrous stage of the cycle secreted an average of 29·5 pg. progesterone cell/day; the cells harvested from the large vascular follicles found at oestrus secreted an average of 173 pg./cell/day. The small, poorly vascularized follicles found adjacent to the large vascular follicles of mares in oestrus yielded cells which secreted less progesterone than those from the larger follicles. Addition of 5 to 10 i.u. human chorionic gonadotrophin (HCG)/ml. at each medium change (every 2–3 days) or for the first 4 days of culture brought about a marked stimulation of progesterone secretion in cultures of ' mid-luteal phase' cells which was maximal after 4 to 7 days. Pregnenolone was converted primarily to progesterone, 20α-hydroxypregn-4-en-3-one and 17-hydroxyprogesterone; the metabolism was not significantly altered by the addition of a mixture of 10 i.u. HCG plus 10 i.u. pregnant mare serum gonadotrophin (PMSG). Cells harvested from mares in oestrus converted pregnenolone to progesterone in a higher yield compared with cells harvested from mares in the midluteal phase of the cycle. Addition of 10 i.u. HCG/ml. or PMSG plus HCG (10 i.u. each/ml.) stimulated aromatization of testosterone by 'midluteal phase' cultures but not by 'oestrous phase' cell cultures. These results demonstrate that the in vivo environment as well as the in vitro conditions influence the steroidogenic activity of equine granulosa cell cultures.

STUDIES ON TISSUE CULTURE OF EQUINE OVARIAN CELL TYPES: STEROIDOGENESIS

1969 ◽  
Vol 43 (3) ◽  
pp. 391-402 ◽  
Author(s):  
CORNELIA P. CHANNING ◽  
SUSAN A. GRIEVES
Keyword(s):  
Tissue Culture ◽  
Granulosa Cell ◽  
Stromal Cells ◽  
Culture Medium ◽  
Horse Serum ◽  
Cell Types ◽  
Gas Liquid Chromatography ◽  
Ovarian Cell ◽  
Constant Rate ◽  
17 Hydroxyprogesterone

SUMMARY Utilizing paper and gas—liquid chromatography, the pattern of steroid secretion by cultures of equine granulosa, thecal and stromal cells has been studied. Granulosa cell cultures secreted primarily progesterone and 20α-hydroxypregn-4-en-3-one. A granulosa cell culture of 700,000 cells harvested from two large vascular follicles of a mare in oestrus secreted the following amounts of steroid from days 0 to 6 of culture: 561 μg. progesterone, 68 μg. 20α-hydroxypregn-4-en-3-one, 5·3 μg. 17-hydroxyprogesterone, 1 μg. androstenedione, 0·12 μg. dehydroepiandrosterone and 2·7 μg. oestradiol. Thecal tissue harvested from the same follicles secreted 29, 3·0, 1·8, 1·4, 1·1 and 0·99 μg., respectively, of the same steroids. All these steroids originated from endogenous precursors or from acetate or cholesterol in the tissue culture medium which contained 15% horse serum, 30% medium '199' and 55% Hanks's solution. Cultures of granulosa cells harvested from two large vascular follicles of a mare in early oestrus secreted progesterone at a constant rate of 35 pg./cell/day between days 6 and 34 of culture, thus demonstrating that these cells do not dedifferentiate in culture. The secretion of 20α-hydroxypregn-4-en-3-one relative to progesterone increased from days 3 to 6 and remained constant from days 6 to 15.

STUDIES ON TISSUE CULTURE OF EQUINE OVARIAN CELL TYPES: PATHWAYS OF STEROIDOGENESIS

1969 ◽  
Vol 43 (3) ◽  
pp. 403-414 ◽  
Author(s):  
CORNELIA P. CHANNING
Keyword(s):  
Tissue Culture ◽  
Granulosa Cell ◽  
Cell Cultures ◽  
Granulosa Cells ◽  
Sodium Acetate ◽  
Steroid Hormone ◽  
Cell Types ◽  
Ovarian Cell ◽  
Luteal Tissue ◽  
17 Hydroxyprogesterone

SUMMARY Equine granulosa cell cultures were incubated with various labelled steroid hormone precursors, and the products of these incubations were purified and characterized by a combination of paper chromatography, derivative formation and recrystallization. Sodium acetate, cholesterol and pregnenolone were converted mainly to progesterone in addition to significant amounts of 20α-hydroxypregn-4-en-3-one and 17-hydroxyprogesterone plus small amounts of dehydroepiandrosterone, androstenedione and oestradiol-17β. Both androstenedione and testosterone were converted to oestrone and oestradiol-17β. Testosterone was converted to androstenedione but the converse did not apply. These experiments demonstrated that equine granulosa cells in culture contain all the enzymes for converting acetate to progesterone and oestradiol-17β and that the pathways are similar to those of luteal tissue.

Tests by Tissue Culture Methods on the Nature of Immunity Transplanted Skin

1948 ◽  
Vol s3-89 (7) ◽  
pp. 239-252
Author(s):  
P. B. MEDAWAR
Keyword(s):  
Tissue Culture ◽  
Cell Division ◽  
Epidermal Cell ◽  
Cell Suspensions ◽  
Acquired Immunity ◽  
Culture Methods ◽  
Division Frequency

The transplantation of skin from one rabbit to another elicits a reaction that conforms in main outline with that of an actively acquired immunity. The experiments described in this paper were designed to test the hypothesis that the regression of such grafts is secured by the action of antibodies demonstrable in vitro. Skin from adult rabbits has therefore been cultivated in the presence of serum and growing mesenchymal tissues derived solely from rabbits heavily and specifically immunized against it. Immune sera and tissues are without effect on the survival, cell-division frequency and migratory activities of explanted skin, and agglutinins for epidermal cell suspensions are not demonstrable in immune sera. With certain stated qualifications, it has therefore been concluded that the occurrence of free antibodies is not a sufficient explanation of the regression of skin homografts in vivo.

STEROIDOGENESIS AND MORPHOLOGY OF HUMAN OVARIAN CELL TYPES IN TISSUE CULTURE

1969 ◽  
Vol 45 (2) ◽  
pp. 297-NP ◽  
Author(s):  
CORNELIA P. CHANNING
Keyword(s):  
Tissue Culture ◽  
Stromal Cells ◽  
Lipid Droplets ◽  
Cell Types ◽  
Growth Characteristics ◽  
Gas Liquid Chromatography ◽  
Culture Cells ◽  
Human Male ◽  
Thecal Cells ◽  
17 Hydroxyprogesterone

SUMMARY Human granulosa, thecal, and stromal cells were grown in tissue culture in a medium containing 15% pooled human male serum, 30% Parker's medium '199', and 55% Hanks's solution. Granulosa cells underwent hyperplasia until about 10–12 days of culture and stayed alive up to at least 100 days. However, from about 20 days onward some of the cells gradually became detached from the glass. Initially the cytoplasm became eosinophilic and granular and accumulated lipid droplets and maintained these characteristics for about 20 days. From day 0–3 of culture, cells from non-gonadotrophin-treated women secreted 10·7 μg. progesterone, 0·54 μg. pregnenolone, 0·52 μg. oestrone and 0·45 μg. oestradiol as determined by gas-liquid chromatography. They also converted pregnenolone to similar products. A pool of thecal cells cultured for 3 days secreted 2·84 μg. progesterone, 0·36 μg. pregnenolone, 1·38 μg. 17-hydroxyprogesterone, 2·10 μg. androstenedione, 1·93 μg. oestrone and 1·44 μg. oestradiol. Stromal cells secreted less steroid per cell and no detectable oestrogens. Thecal cells grew in stellate colonies and had a lacy cytoplasm which contained vacuoles of various sizes which did not contain large amounts of lipid. Stromal cells resembled fibroblasts in morphology and growth characteristics.

scholarly journals EEA1, a Tethering Protein of the Early Sorting Endosome, Shows a Polarized Distribution in Hippocampal Neurons, Epithelial Cells, and Fibroblasts

2000 ◽  
Vol 11 (8) ◽  
pp. 2657-2671 ◽  
Author(s):  
Jean M. Wilson ◽  
Meltsje de Hoop ◽  
Natasha Zorzi ◽  
Ban-Hock Toh ◽  
Carlos G. Dotti ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mammalian Cells ◽  
Hippocampal Neurons ◽  
Cell Types ◽  
Early Endosomes ◽  
Filamentous Material ◽  
Endocytic Vesicles ◽  
Fibroblastic Cells ◽  
Darby Canine Kidney ◽  
Endocytic Pathways

EEA1 is an early endosomal Rab5 effector protein that has been implicated in the docking of incoming endocytic vesicles before fusion with early endosomes. Because of the presence of complex endosomal pathways in polarized and nonpolarized cells, we have examined the distribution of EEA1 in diverse cell types. Ultrastructural analysis demonstrates that EEA1 is present on a subdomain of the early sorting endosome but not on clathrin-coated vesicles, consistent with a role in providing directionality to early endosomal fusion. Furthermore, EEA1 is associated with filamentous material that extends from the cytoplasmic surface of the endosomal domain, which is also consistent with a tethering/docking role for EEA1. In polarized cells (Madin-Darby canine kidney cells and hippocampal neurons), EEA1 is present on a subset of “basolateral-type” endosomal compartments, suggesting that EEA1 regulates specific endocytic pathways. In both epithelial cells and fibroblastic cells, EEA1 and a transfected apical endosomal marker, endotubin, label distinct endosomal populations. Hence, there are at least two distinct sets of early endosomes in polarized and nonpolarized mammalian cells. EEA1 could provide specificity and directionality to fusion events occurring in a subset of these endosomes in polarized and nonpolarized cells.

scholarly journals Regulation of Division and Differentiation of Plant Stem Cells

2018 ◽  
Vol 34 (1) ◽  
pp. 289-310 ◽  
Author(s):  
Edith Pierre-Jerome ◽  
Colleen Drapek ◽  
Philip N. Benfey
Keyword(s):  
Stem Cell ◽  
Cell Division ◽  
Gene Regulatory Networks ◽  
Regulatory Networks ◽  
Cell Types ◽  
Stem Cell Maintenance ◽  
Cell Position ◽  
Dependent Cell ◽  
Gene Regulatory ◽  
Plant Stem

A major challenge in developmental biology is unraveling the precise regulation of plant stem cell maintenance and the transition to a fully differentiated cell. In this review, we highlight major themes coordinating the acquisition of cell identity and subsequent differentiation in plants. Plant cells are immobile and establish position-dependent cell lineages that rely heavily on external cues. Central players are the hormones auxin and cytokinin, which balance cell division and differentiation during organogenesis. Transcription factors and miRNAs, many of which are mobile in plants, establish gene regulatory networks that communicate cell position and fate. Small peptide signaling also provides positional cues as new cell types emerge from stem cell division and progress through differentiation. These pathways recruit similar players for patterning different organs, emphasizing the modular nature of gene regulatory networks. Finally, we speculate on the outstanding questions in the field and discuss how they may be addressed by emerging technologies.

Cultured epithelial cells derived from human foetal pancreas as a model for the study of cystic fibrosis: further analyses on the origins and nature of the cell types

1988 ◽  
Vol 90 (1) ◽  
pp. 73-77
Author(s):  
A. Harris ◽  
L. Coleman
Keyword(s):  
Cystic Fibrosis ◽  
Tissue Culture ◽  
Epithelial Cells ◽  
Culture System ◽  
Cultured Cells ◽  
Cell Types ◽  
Cultured Epithelial Cells ◽  
Different Cell Types ◽  
Foetal Pancreas

The establishment of a tissue-culture system for epithelial cells derived from human foetal pancreas has recently been reported. Further analyses have now been made on these cells in vitro, together with parallel investigation of the distribution of different cell types within the intact foetal pancreas. Results support the view that the cultured cells are ductal in origin and nature. Pancreatic epithelial cell cultures have also been established from foetuses with cystic fibrosis.

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