Culture of equine trophoblastic vesicles in vitro.
Abstract: Trophoblastic vesicles have been used to study early embryonic development and maternal recognition of pregnancy in domestic animals. The purpose of this study was to characterize the formation of trophoblastic vesicles from Day-12 to Day-16 equine conceptuses. Conceptuses (n = 19) were collected nonsurgically from mares, the capsule was removed, and the conceptus (trophoblast and inner cell mass) was dissected into 2- to 4-mm fragments. Conceptus fragments were cultured in either Ham's F10 (HF10) or Minimum Essential Media (MEM) with 10% fetal bovine serum (FBS) in 24-well plates. Plates were incubated at 37 degrees C in a humidified atmosphere of 90% N(2), 5% O(2), and 5% CO(2) and were examined at 48 and 96 h for the number and diameter of trophoblastic vesicles formed. There was no significant difference (P > 0.1) between HF10 and MEM in the diameter of trophoblastic vesicles at 48 or 96 h of culture. There was, however, a significant increase (P < 0.01) in the number of trophoblastic vesicles per conceptus between 48 and 96 h of culture for HF10 and MEM. The mean diameters of trophoblastic vesicles after 96 h in culture were 396.4 +/- 19.9 mum and 415.0 +/- 12.0 mum for HF10 and MEM, respectively. Histologic sections of trophoblastic vesicles revealed a bilaminar structure consistent with the presence of trophectoderm and endoderm. Squash preparations of trophoblastic vesicles contained cells similar to those recognized in squash preparations of fresh conceptuses. Areas of increased cell density that resembled the inner-cell mass were seen in both fresh trophoblastic vesicles and in sections of trophoblastic vesicles. Equine trophoblastic vesicles may be useful to further the study of early embryonic development and pregnancy recognition in mares.
Publication Date: 1989-09-01 PubMed ID: 16726686DOI: 10.1016/0093-691x(89)90006-xGoogle Scholar: Lookup
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- Journal Article
Summary
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The research paper mainly focuses on the study of trophoblastic vesicles in horses to better understand early embryonic development and pregnancy recognition. The use of conceptuses in a laboratory environment enabled the researchers to observe the formation of these vesicles under controlled conditions, with an interest in understanding their role.
Methodology
- Conceptuses (early stage embryos) were collected non-surgically from mares and dissected into small fragments.
- These fragments were then placed into culture in either Ham’s F10 (HF10) or Minimum Essential Media (MEM) with 10% fetal bovine serum (FBS) which are common mediums used in cell culture.
- The cultures were kept in 24-well plates and were incubated at 37 degrees Celsius in a special atmosphere consisting of 90% Nitrogen, 5% Oxygen, and 5% Carbon Dioxide.
- The researchers then examined these cultures at 48 and 96 hours to count the number of trophoblastic vesicles that had formed and measure their diameter.
Findings
- No significant difference was found between the two types of media (HF10 and MEM) in terms of the diameter of the trophoblastic vesicles that formed at either 48 or 96 hours of culture.
- There was, however, a significant increase in the number of trophoblastic vesicles per conceptus between 48 and 96 hours of culture for both types of media.
- The vesicles had a bilaminar structure consistent with the presence of trophectoderm and endoderm. These are the two types of cells that typically make up these vesicles in a developing embryo.
- Cells similar to those found in fresh (non-cultured) conceptuses were found in the cultured vesicles.
Conclusion
- The results indicate that the equine trophoblastic vesicles can be used to study and expand our knowledge about early embryonic development and the recognition of pregnancy in mares.
- The ability to cultivate these vesicles in a laboratory setting and the relevance of the vesicles to embryonic development and pregnancy recognition indicates their potential use as a tool for further research.
Cite This Article
APA
Ball BA, Altschul M, Freeman KP, Hillman RB.
(1989).
Culture of equine trophoblastic vesicles in vitro.
Theriogenology, 32(3), 401-412.
https://doi.org/10.1016/0093-691x(89)90006-x Publication
Researcher Affiliations
- Department of Clinical Sciences Cornell University Ithaca, NY 14853 USA.
Citations
This article has been cited 1 times.- Swegen A. Maternal recognition of pregnancy in the mare: does it exist and why do we care?. Reproduction 2021 May 5;161(6):R139-R155.
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