A Method of Growing Chick Embryos In Vitro

Boone, M. A.

doi:10.3382/ps.0420916

Cited by 11 publications

(4 citation statements)

References 3 publications

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“…In the past, investigators have cultured embryos in glass dishes (Assheton, 1896;Fere, 1900;Loisel, 1900;Paton, 1911;Vogelaar and van den Boogert, 1925;Vollmar, 1935;Schmidt, 19371, ceramic dishes (Corner and Richter, 1973), beakers (Romanoff, 1943;Boone, 1963;Palen and Thorneby, 1976), plastic shells (Quisenberry and Dillon, 1962), petri dishes (Williams and Boone, 1961), sandwich bags (Schlesinger, 1966;Elliott and Bennett, 1971), and in sandwich-type plastic wrap in several configurations (Criley, 1971;Dunn, 1974) with varying degrees of success. Only relatively recently, however, has shell-less culture become a useful laboratory technique.…”

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confidence: 99%

Effects of varying chamber construction and embryo pre‐incubation age on survival and growth of chick embryos in shell‐less culture

1981

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Shell-less culture involves culturing chick embryos with associated yolk and albumen outside of the eggshell and shell membranes. The technique allows direct access to and continuous observation of cultured embryos almost to the time of hatching. The plastic wrap/culture tripod technique described in this paper allows normal embryonic growth and differentiation from 48 hours (in ovo pre-incubation age) through at least 10 days of total incubation. As the duration of in ovo pre-incubation is decreased below 36 hours, there is a concomitant increase in the percentage of grossly abnormal embryos associated with decreased survival and retarded growth and differentiation. Survival of embryos pre-incubated for 72 hours through 13 and 18 days of total incubation is greater than 80% and 40%, respectively. The wrap/tripod technique allows substantially better survival and more normal development of cultured embryos than does the petri dish technique of Auerbach et al. (1974). Embryonic growth in 7.8-cm-diameter chambers is significantly greater than in either larger (10.6 cm) or smaller (5.2 cm) diameter chambers. Some gas exchange through the culture chamber walls appears necessary for optimal embryonic survival and growth. Suspending egg contents in either Safeway or Handi-Wrap plastic wrap (both of intermediate permeability) in 7.8-cm tripods resulted in superior growth and/or survival compared to suspension in Silastic sheeting (high permeability), Saran Wrap (low permeability), glass dishes (nonpermeable), or glass dishes lined with an inner layer of Safeway wrap(nonpermeable).

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confidence: 99%

Effects of varying chamber construction and embryo pre‐incubation age on survival and growth of chick embryos in shell‐less culture

1981

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“…To the knowledge of the present investigator, this is the first description of a method for culturing embryos from the blastoderm stage through to hatching in a shell‐less culture environment. In previous studies by multiple investigators, drying out of the blastoderm and yolk surface severely limited the success of shell‐less culture of previously unincubated egg contents (Boone, 1963; Romanoff, 1943; Vogelaar & van den Boogert, 1925). In ovo, the blastoderm secretes fluid downward from the overlying albumen into the underlying yolk in chicken (New, 1956) and duck (van Deth, 1962) eggs.…”

Section: Discussionmentioning

confidence: 99%

“…Early authors attempting to maintain previously unincubated egg contents in shell‐less culture reported that the uppermost surface of the yolk frequently appeared “dried out” after a short period of incubation despite high humidity (Boone, 1963; Romanoff, 1943; Vogelaar & van den Boogert, 1925). In 1960, P. H. S. Silver described a technique for the prevention of yolk drying in early chicken egg contents undergoing surgical manipulation of the vitelline membrane in ovo.…”

Section: Introductionmentioning

confidence: 99%

Shell‐less culture system for chick embryos from the blastoderm stage to hatching

Dunn

2023

J Exp Zool Pt A

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Since 2014, methods have been described to hatch chick embryos from shell‐less culture after egg contents are first incubated within shells for 55–70 h. The present report describes for the first time a shell‐less culture system for chick embryos from the blastoderm stage to hatching. For the first 69–70 h, egg contents suspended in polymethylpentene kitchen wrap (F.O.R. Wrap, Riken Fabro, Tokyo, Japan) supported in 6.35 or 6.67 cm inside diameter tripods and covered with a disc of immobilized Milli‐Wrap, were rotated back and forth through 90° at 16 or 22 cycles per minute (CPM). Subsequently, the Milli‐Wrap disc was removed and culture tripods were transferred to environmental chambers, which were rocked ±20° through incubation day 8.5 (E8.5). From E9, environmental chambers were maintained in the horizontal position through to hatching with controlled O2 and CO2. To provide supplemental calcium, an aqueous solution containing 100 mg/mL of calcium l‐lactate hydrate was injected through the plastic wrap into the albumen at E9 (2.5 mL) and at E13 (1.0 mL) or E15 (1.0 mL). After incubation for 69–70 h at 16 or 22 CPM, 80%–83% of previously unincubated egg contents yielded apparently normal embryos. Hatch rate of normal embryos resulting from turntable incubation at 16 or 22 CPM was approximately 43%. Of note, egg contents remained in the same culture tripod from blastoderm stage to hatching. This technique may find use as an educational tool and in basic investigations of early embryogenesis, teratogenesis, and gene transfer experiments.

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“…Ex-ovo embryonic development is the extraction of the embryo from the eggshell, which is placed in a medium that will provide the necessary conditions for its development, thereby helping the observation and manipulation of the organism (Kamihira et al, 2005). In the studies performed by Boone (1963), the embryos were incubated in 50 and 100 mL beakers, and the different materials that could provide better development for the organisms were examined. It was found that the use of 100 mL beakers favored better development than the use of 50 mL beakers, and the use of a plastic wrap halted the humidity loss and provided the right gas exchange.…”

Section: Introductionmentioning

confidence: 99%

Study of ex-ovo Embryonic Development of Gallus gallus domesticus

Macías-Marín,

Guerrero-Barrera,

Valdivia-Flores

et al. 2023

Int. J. Morphol.

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The domestic chicken is a species of bird that has been extensively studied in regard to its biology and as a model organism for science. The reproduction of the species is by the laying of fertilized eggs, which in a period of 21 days will develop a chick inside. Several methods have been described to develop embryos ex-ovo, allowing the observation and manipulation of the organism. This work has the propose to standardize a method that allows the development of the embryos inside the artificial incubation system, which has a low cost and is easy to make. In this work, 100 chicken eggs were used to study the effects of humidity, mineral supplementation, and the preincubation time of the egg on the incubation ex-ovo of the embryos. Embryo development was documented through the different days. Pulverized eggshell was selected as an optimal source to provide calcium, magnesium, phosphorus, and other minerals to the developing embryo. By providing 900-1200 mg of pulverized eggshell, 40 mL of the 0.001 % solution of benzalkonium chloride, and a preincubation time of approximately 56 h, the embryos were able to develop until 19 days, and even though they did not reach hatching, the incubation conditions that allowed the survival and development of embryos until late stages were achieved. Thus, due to the conditions established for calcium, humidity and preincubation time, in the present work, the chicks reached 19 days of development.

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A Method of Growing Chick Embryos In Vitro

Cited by 11 publications

References 3 publications

Effects of varying chamber construction and embryo pre‐incubation age on survival and growth of chick embryos in shell‐less culture

Effects of varying chamber construction and embryo pre‐incubation age on survival and growth of chick embryos in shell‐less culture

Shell‐less culture system for chick embryos from the blastoderm stage to hatching

Study of ex-ovo Embryonic Development of Gallus gallus domesticus

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