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Determination

Determination

During embryogenesis, cells become progressively committed to different fates through a two-step process: specification followed by determination. Specification is demonstrated by removing a segment of an early embryo, “neutrally” culturing the tissue in vitro—for example, in a petri dish with simple medium—and then observing the derivatives. If the cultured region gives rise to cell types that it would normally generate in the embryo, this means that it is specified. In contrast, determination...

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Related Experiment Video

Updated: Jun 10, 2026

Preparation of Rat Serum Suitable for Mammalian Whole Embryo Culture

Preparation of Rat Serum Suitable for Mammalian Whole Embryo Culture

Published on: August 3, 2014

Complement component C3 functions as an embryotrophic factor in early postimplantation rat embryos.

Makoto Usami¹, Katsuyoshi Mitsunaga, Atsuko Miyajima

¹Division of Pharmacology, National Institute of Health Sciences, Tokyo, Japan. usami@nihs.go.jp

The International Journal of Developmental Biology

|August 17, 2010

Summary

Complement component C3 (C3) acts as an embryotrophic factor for early rat embryo development. This protein supports embryonic growth through novel mechanisms, likely involving the visceral yolk sac.

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Preparation of Rat Serum Suitable for Mammalian Whole Embryo Culture

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Published on: August 3, 2014

Embryo Microinjection and Electroporation in the Chordate Ciona intestinalis

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The Method of Rodent Whole Embryo Culture using the Rotator-type Bottle Culture System

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Published on: August 28, 2010

Area of Science:

Reproductive Biology
Immunology
Developmental Biology

Background:

Early postimplantation embryonic development relies on nutrient supply.
The complement system, particularly complement component C3 (C3), has been traditionally viewed as potentially harmful to embryos.

Purpose of the Study:

To identify the embryotrophic factor in rat serum.
To investigate the role of complement component C3 (C3) in early embryonic development.
To elucidate the mechanism of C3's embryotrophic activity.

Main Methods:

Partial purification of rat serum to isolate an embryotrophic factor.
N-terminal sequence analysis to identify the factor as C3.
In vitro culture of rat embryos with purified rat and rabbit C3.
C3-depletion experiments using cobra venom factor.
Immunochemical analysis and fluorescence labeling to determine C3 localization.

Main Results:

Partially purified rat serum identified C3 as an embryotrophic factor.
Purified rat C3 supported rat embryo growth in vitro, with optimal activity at 0.5 mg/ml.
Rat embryos selectively consumed C3 in culture; C3 depletion impaired growth.
Methylamine treatment did not affect C3's embryotrophic activity, indicating the thiolester bond is not critical.
Rabbit C3 showed weak activity, suggesting species specificity.
C3 was specifically localized to the visceral yolk sac, not the embryo proper.
Unfragmented C3 bound more strongly to the visceral yolk sac than C3b.

Conclusions:

Complement component C3 (C3) functions as an embryotrophic factor in early postimplantation rat development.
C3 likely exerts its supportive role through the visceral yolk sac via novel mechanisms.
This finding challenges the traditional view of C3 as solely detrimental and offers new insights into embryonic growth and C3 function.