scholarly journals Effects of growth factors during in vitro culture of mouse and rat embryos

2015 ◽  
Vol 19 (4) ◽  
pp. 372
Author(s):  
E. Yu. Brusentsev ◽  
T. N. Igonina ◽  
I. N. Rozhkova ◽  
D. S. Ragaeva ◽  
S. Ya. Amstislavsky
Keyword(s):  
Growth Factors ◽  
In Vitro Culture ◽  
Rat Embryos

Effects of growth factors during in vitro culture of mouse and rat embryos

2016 ◽  
Vol 6 (4) ◽  
pp. 378-383 ◽  
Author(s):  
E. Yu. Brusentsev ◽  
T. N. Igonina ◽  
I. N. Rozhkova ◽  
D. S. Ragaeva ◽  
S. Ya. Amstislavsky
Keyword(s):  
Growth Factors ◽  
In Vitro Culture ◽  
Rat Embryos

In vitro Culture of the Eye and the Retina of the Mouse and Rat

Development ◽  
1958 ◽  
Vol 6 (1) ◽  
pp. 178-182
Author(s):  
D. R. Lucas ◽  
O. A. Trowell
Keyword(s):  
In Vitro Culture ◽  
Rosette Formation ◽  
Normal Rate ◽  
Embryonic Chick ◽  
Prominent Feature ◽  
Rat Embryos ◽  
Embryo Extract ◽  
Isolated Retina

Strange ways & Fell (1926) and Dorris (1938) showed that embryonic chick eyes could be cultivated successfully in vitro on plasma clots, and Harrison (1951) and Reinbold (1954) obtained satisfactory results with an agar-tyrode medium containing embryo extract. The retina in these eyes was undifferentiated and, though little or no increase in the size of the eyes occurred, the authors agreed that cytological differentiation proceeded at a more or less normal rate. Tansley (1933) reported the successful cultivation on plasma clots of whole eyes from rat embryos of 14–17 days and of isolated retina from rats up to 5 days old. Cytological differentiation proceeded at a normal rate, at least for the first few days in culture, but excessive growth of mesodermal elements tended to occur after 12 to 14 days. Rosette formation was a prominent feature in these cultures.

In vitro differentiation of theca cells from ovarian cells isolated from postmenopausal women

2020 ◽  
Vol 35 (12) ◽  
pp. 2793-2807
Author(s):  
P Asiabi ◽  
M M Dolmans ◽  
J Ambroise ◽  
A Camboni ◽  
C A Amorim
Keyword(s):  
Growth Factors ◽  
In Vitro Culture ◽  
Protein Detection ◽  
Basic Medium ◽  
Specific Marker ◽  
Mrna Levels ◽  
Theca Cells ◽  
Ovarian Cortex ◽  
Steroid Dehydrogenase

Abstract STUDY QUESTION Can human theca cells (TCs) be differentiated in vitro? SUMMARY ANSWER It is possible to differentiate human TCs in vitro using a medium supplemented with growth factors and hormones. WHAT IS KNOWN ALREADY There are very few studies on the origin of TCs in mammalian ovaries. Precursor TCs have been described in neonatal mice ovaries, which can differentiate into TCs under the influence of factors from oocytes and granulosa cells (GCs). On the other hand, studies in large animal models have reported that stromal cells (SCs) isolated from the cortical ovarian layer can also differentiate into TCs. STUDY DESIGN, SIZE, DURATION After obtaining informed consent, ovarian biopsies were taken from eight menopausal women (53–74 years of age) undergoing laparoscopic surgery for gynecologic disease not related to the ovaries. SCs were isolated from the ovarian cortex and in vitro cultured for 8 days in basic medium (BM) (G1), enriched with growth factors, FSH and LH in plastic (G2) or collagen substrate without (G3) or with (G4) a GC line. PARTICIPANTS/MATERIALS, SETTING, METHODS To confirm TC differentiation, relative mRNA levels for LH receptor (Lhr), steroidogenic acute regulatory protein (Star), cholesterol side-chain cleavage enzyme (Cyp11a1), cytochrome P450 17A1 (Cyp17a1), hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 (Hsd3b1) and hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 2 (Hsd3b2) were assessed. Immunohistochemistry was also performed for their protein detection and a specific marker was identified for TCs (aminopeptidase-N, CD13), as were markers for theca and small luteal cells (dipeptidyl peptidase IV (CD26) and Notch homolog 1, translocation-associated (NOTCH1)). Finally, we analyzed cell ultrastructure before (Day 0) and after in vitro culture (Day 8), and dehydroepiandrosterone (DHEA) and progesterone levels in the medium using transmission electron microscopy (TEM) and ELISA, respectively. MAIN RESULTS AND THE ROLE OF CHANCE Results obtained from qPCR showed a significant increase (P < 0.05) in mRNA levels of Lhr in F2 (floating cells in G2) and G4, Cyp17a1 in G1 and F1 (floating cells in G1) and Hsd3b2 in G1, G2, G3 and G4. Immunohistochemistry confirmed expression of each enzyme involved in the steroidogenic pathway at the protein stage. However, apart from G1, all other groups exhibited a significant (P < 0.05) rise in the number of CD13-positive cells. There was also a significant increase (P < 0.05) in NOTCH1-positive cells in G3 and G4. Ultrastructure analyses by TEM showed a distinct difference between groups and also versus Day 0. A linear trend with time revealed a significant gain (q < 0.001) in DHEA concentrations in the medium during the culture period in G1, G2, G3 and G4. It also demonstrated a statistical increase (q < 0.001) in G2, G3 and G4 groups, but G1 remained the same throughout culture in terms of progesterone levels. LARGE SCALE DATA N/A LIMITATIONS, REASONS FOR CAUTION Shorter periods of in vitro culture (e.g. 2, 4 and 6 days) could have led to increased concentrations of differentiated TCs in G2, G3 and G4. In addition, a group of cells cultured in BM and accompanied by COV434 cells would be necessary to understand their role in the differentiation process. Finally, while our results demonstrate that TCs can be differentiated in vitro from cells isolated from the cortical layer of postmenopausal ovaries, we do not know if these cells are differentiated from a subpopulation of precursor TCs present in ovarian cortex or ovarian SCs in general. It is therefore necessary to identify specific markers for precursor TCs in human ovaries to understand the origin of these cells. WIDER IMPLICATIONS OF THE FINDINGS This is a promising step toward understanding TC ontogenesis in the human ovary. Moreover, in vitro-generated human TCs can be used for studies on drug screening, as well as to understand TC-associated pathologies, such as androgen-secreting tumors and polycystic ovary syndrome. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (FNRS) (C.A.A. is an FRS-FNRS Research Associate; grant MIS #F4535 16 awarded to C.A.A.; grant 5/4/150/5 awarded to M.M.D.; grant ASP-RE314 awarded to P.A.) and Foundation Against Cancer (grant 2018-042 awarded to A.C.). The authors declare no competing interests.

In Vitro Culture of Chondrocytes in a Novel Thermoreversible Gelation Polymer Scaffold Containing Growth Factors

2006 ◽  
Vol 12 (5) ◽  
pp. 1237-1245 ◽  
Author(s):  
Ayuko Yasuda ◽  
Koji Kojima ◽  
Kevin W. Tinsley ◽  
Hiroshi Yoshioka ◽  
Yuichi Mori ◽  
...  
Keyword(s):  
Growth Factors ◽  
In Vitro Culture ◽  
Polymer Scaffold ◽  
Thermoreversible Gelation ◽  
Thermoreversible Gelation Polymer

Growth factors in invertebrate in vitro culture

1991 ◽  
Vol 27 (6) ◽  
pp. 483-486 ◽  
Author(s):  
Stephen M. Ferkovich ◽  
Herbert Oberlander
Keyword(s):  
Growth Factors ◽  
In Vitro Culture

In vitro culture of bovine IVM-IVF embryos: Cooperative interaction among embryos and the role of growth factors

1994 ◽  
Vol 41 (6) ◽  
pp. 1323-1331 ◽  
Author(s):  
C.L. Keefer ◽  
S.L. Stice ◽  
A.M. Paprocki ◽  
P. Golueke
Keyword(s):  
Growth Factors ◽  
In Vitro Culture ◽  
Cooperative Interaction

Transplantation of metanephroi after preservation in vitro

2001 ◽  
Vol 281 (2) ◽  
pp. R661-R665 ◽  
Author(s):  
Sharon A. Rogers ◽  
Marc R. Hammerman
Keyword(s):  
Growth Factors ◽  
Normal Structure ◽  
Sprague Dawley ◽  
Adult Rats ◽  
Rat Embryos ◽  
University Of Wisconsin ◽  
Distal Tubules ◽  
And Function ◽  
First Time

To determine whether transplanted metanephroi grow, differentiate, and function in hosts after preservation in vitro, we implanted metanephroi from embryonic day 15 ( E15) Sprague-Dawley rat embryos into the omentum of nonimmunosuppressed uninephrectomized Sprague-Dawley (host) rats. Metanephroi were either implanted directly or suspended in ice-cold University of Wisconsin (UW) preservation solution with or without added growth factors for 3 days before implantation. The size and extent of tissue differentiation preimplantation of E15 metanephroi implanted directly were not distinguishable from the size and differentiation of metanephroi preserved for 3 days. In contrast, E16 metanephroi were larger than E15 metanephroi preserved for 3 days. E16 metanephroi or E13 metanephroi grown in organ culture for 3 days contained more differentiated nephron structures than those in E15 metanephroi preserved for 3 days. By 4 wk posttransplantation, metanephroi that had been preserved for 3 days had grown and differentiated such that glomeruli, proximal and distal tubules, and collecting ducts with normal structure had developed. At 12 wk posttransplantation, inulin clearances of preserved metanephroi were comparable to those of metanephroi that had been implanted directly. Addition of growth factors to the UW solution enhanced inulin clearances. Here we show for the first time that functional kidneys develop from metanephroi transplanted from rat embryos to adult rats after as long as 3 days of preservation in vitro.

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