Successful in vitro growth of rat two-cell embryos to blastocysts using a simple chemically defined medium

V. H.H. Goh, S. K. Adiga, C. F. Tain, T. Y.Y. Tong, S. C. Ng

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The aim of the present study was to formulate a simple chemically defined medium for the in vitro growth of rat two-cell embryos to blastocysts. Embryos from day 2 pregnant rats were retrieved and placed in paraffin oil-covered droplets of "rat two-cell embryo culture medium" (R2ECM) containing combinations of various serum supplements, glucose, L-glutamine, and cultured up to 96 h in a CO2 incubator. Embryos cultured in the basic medium (R2ECM), as well as those supplemented either with fetal bovine serum (FBS) or male rat serum (MRS) did not develop beyond the two- to four-cell stage. In R2ECM with 0.3% bovine serum albumin (BSA) and 7.5 mM glucose, 44% of embryos reached the blastocyst stage by 96 h in culture, and the blastulation rate increased to about 83% when 1 mM of L-glutamine was added. To evaluate the effects of varying doses of glucose, two-cell embryos were cultured in R2ECM supplemented with 0.3% BSA, 1 mM L-glutamine, and 2.5, 5.0, or 7.5 mM of glucose. The percentage of embryos reaching the blastocyst stage for 2.5, 5.0, and 7.5 mM glucose was 64.6%, 65.3%, and 82.9%, respectively. The present study showed that the modified medium (R2ECM) is a simple chemically defined medium that is capable of supporting in vitro growth of rat two-cell embryos to blastocysts in high proportion (greater than 80%) without the need for change of medium within 96 h of culture.

Original languageEnglish
Pages (from-to)171-175
Number of pages5
JournalJournal of Pharmacological and Toxicological Methods
Volume43
Issue number3
DOIs
Publication statusPublished - 2000

Fingerprint

Blastocyst
Rats
Embryonic Structures
Glucose
Glutamine
Growth
Bovine Serum Albumin
Cell culture
Serum
Culture Media
Incubators
In Vitro Techniques
Cell Culture Techniques

All Science Journal Classification (ASJC) codes

  • Toxicology
  • Pharmacology

Cite this

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title = "Successful in vitro growth of rat two-cell embryos to blastocysts using a simple chemically defined medium",
abstract = "The aim of the present study was to formulate a simple chemically defined medium for the in vitro growth of rat two-cell embryos to blastocysts. Embryos from day 2 pregnant rats were retrieved and placed in paraffin oil-covered droplets of {"}rat two-cell embryo culture medium{"} (R2ECM) containing combinations of various serum supplements, glucose, L-glutamine, and cultured up to 96 h in a CO2 incubator. Embryos cultured in the basic medium (R2ECM), as well as those supplemented either with fetal bovine serum (FBS) or male rat serum (MRS) did not develop beyond the two- to four-cell stage. In R2ECM with 0.3{\%} bovine serum albumin (BSA) and 7.5 mM glucose, 44{\%} of embryos reached the blastocyst stage by 96 h in culture, and the blastulation rate increased to about 83{\%} when 1 mM of L-glutamine was added. To evaluate the effects of varying doses of glucose, two-cell embryos were cultured in R2ECM supplemented with 0.3{\%} BSA, 1 mM L-glutamine, and 2.5, 5.0, or 7.5 mM of glucose. The percentage of embryos reaching the blastocyst stage for 2.5, 5.0, and 7.5 mM glucose was 64.6{\%}, 65.3{\%}, and 82.9{\%}, respectively. The present study showed that the modified medium (R2ECM) is a simple chemically defined medium that is capable of supporting in vitro growth of rat two-cell embryos to blastocysts in high proportion (greater than 80{\%}) without the need for change of medium within 96 h of culture.",
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Successful in vitro growth of rat two-cell embryos to blastocysts using a simple chemically defined medium. / Goh, V. H.H.; Adiga, S. K.; Tain, C. F.; Tong, T. Y.Y.; Ng, S. C.

In: Journal of Pharmacological and Toxicological Methods, Vol. 43, No. 3, 2000, p. 171-175.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Successful in vitro growth of rat two-cell embryos to blastocysts using a simple chemically defined medium

AU - Goh, V. H.H.

AU - Adiga, S. K.

AU - Tain, C. F.

AU - Tong, T. Y.Y.

AU - Ng, S. C.

PY - 2000

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AB - The aim of the present study was to formulate a simple chemically defined medium for the in vitro growth of rat two-cell embryos to blastocysts. Embryos from day 2 pregnant rats were retrieved and placed in paraffin oil-covered droplets of "rat two-cell embryo culture medium" (R2ECM) containing combinations of various serum supplements, glucose, L-glutamine, and cultured up to 96 h in a CO2 incubator. Embryos cultured in the basic medium (R2ECM), as well as those supplemented either with fetal bovine serum (FBS) or male rat serum (MRS) did not develop beyond the two- to four-cell stage. In R2ECM with 0.3% bovine serum albumin (BSA) and 7.5 mM glucose, 44% of embryos reached the blastocyst stage by 96 h in culture, and the blastulation rate increased to about 83% when 1 mM of L-glutamine was added. To evaluate the effects of varying doses of glucose, two-cell embryos were cultured in R2ECM supplemented with 0.3% BSA, 1 mM L-glutamine, and 2.5, 5.0, or 7.5 mM of glucose. The percentage of embryos reaching the blastocyst stage for 2.5, 5.0, and 7.5 mM glucose was 64.6%, 65.3%, and 82.9%, respectively. The present study showed that the modified medium (R2ECM) is a simple chemically defined medium that is capable of supporting in vitro growth of rat two-cell embryos to blastocysts in high proportion (greater than 80%) without the need for change of medium within 96 h of culture.

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JO - Journal of Pharmacological and Toxicological Methods

JF - Journal of Pharmacological and Toxicological Methods

SN - 1056-8719

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