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Biomedical subjects

M T Kane

Publications and source records attributed to M T Kane.

At least 55 records · Page 3Linked to original sources

A low molecular weight extract of bovine serum albumin stimulates rabbit blastocyst cell division and expansion in vitro.

Fraction V bovine serum albumin (BSA) was dissolved in 5% formic acid and filtered through a molecular filter with a cutoff of Mr 10 000. The freeze-dried filtrate stimulated increased cell division of cultured rabbit morulae to blastocysts (up to 8-fold increase in cell number) and increased blastocyst expansion (greater than 2-fold). These results show that some samples of commercial BSA contain an embryonic growth factor of a low molecular weight.

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Evidence that protease action is not specifically involved in the hatching of rabbit blastocysts caused by commercial bovine serum albumin in culture.

Commercial samples of bovine serum albumin (BSA) in a complex medium caused growth of 1-cell rabbit embryos to completely hatched blastocysts. Heat treatment of the BSA at 65 or 80 degrees C significantly decreased blastocyst formation and expansion and destroyed the ability to cause blastocyst hatching. Addition of trypsin at levels down to 20 ng/ml caused the formation of hatched blastocysts which degenerated rapidly. The effects of 5 protease inhibitors (ovomucoid trypsin inhibitor, alpha-1-antitrypsin, TAME, TLCK and soybean) were tested. Ovomucoid trypsin inhibitor, TAME and TLCK significantly inhibited blastocyst hatching but only at the highest concentration used. These inhibitors also reduced blastocyst formation and expansion, indicating that their effect was not specifically on blastocyst hatching in vitro. It is concluded that hatching of rabbit blastocysts is probably not dependent on protease action.

Animals↗

Variability in different lots of commercial bovine serum albumin affects cell multiplication and hatching of rabbit blastocysts in culture.

Rabbit morulae were cultured in vitro for 4 days in a synthetic culture medium supplemented with two different lots of commercial bovine serum albumin (BSA) and two different amino acid formulations in a factorial 2 x 2 arrangement. One lot of BSA caused complete hatching of a proportion of blastocysts and formation of more than twice as many cells per blastocyst (hatched and unhatched) as that of the second BSA lot which did not cause complete hatching of any blastocysts. The mean cell numbers of hatched blastocysts were more than twice those of non-hatched blastocysts. There was no significant effect of amino acid formulation.

Animals↗

The role of commercial bovine serum albumin preparations in the culture of one-cell rabbit embryos to blastocysts.

Normal bovine serum albumin (BSA) in a complete medium without energy substrates promoted growth of 1-cell embryos to hatched blastocysts. Defatted charcoal-treated BSA did not promote growth to the blastocyst stage but the addition of pyruvate or palmitic and oleic acids allowed blastocyst growth but not blastocyst hatching. Sodium dodecyl sulphate-gel electrophoresis showed that both the normal and defatted BSA samples were heavily contaminated by proteins other than albumin. Fractionation of the normal BSA on Sephadex G-200 indicated that the property of promoting complete blastocyst hatching was not due to the albumin but was associated with the higher mol. wt fraction of the BSA. Extraction of normal BSA with chloroform appeared to destroy the hatching-promoting ability as neither the residue after extraction nor defatted BSA to which the organic extractate had been added promoted complete blastocyst hatching. It is concluded that commercial BSA may have at least two effects on blastocyst growth: (1) energy provision via albumin-bound fatty acids, and (2) promotion of blastocyst hatching by a non-albumin component.

Animals↗

Measurement theory.

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Clinical Competence↗

The effects of inhibitors of energy metabolism on the growth of one-cell rabbit ova to blastocysts in vitro.

Fertilized 1-cell rabbit ova were cultured in the presence of three oxidative phosphorylation inhibitors (cyanide, 2,4-dinitrophenol and oligomycin), two tricarboxylic acid (TCA) cycle inhibitors (malonate and fluoroacetate) and one glycolytic inhibitor (2-deoxyglucose). All three oxidative phosphorylation inhibitors killed ova at the 1-cell stage and the damage caused by each was similar. Malonate was non-toxic at all concentrations whereas some concentrations of fluoroacetate stopped growth at the 1-cell stage. This toxic effect could, in some circumstances, be reversed by the presence of acetate but not of glucose. 2-Deoxyglucose blocked only the transition from morula to blastocyst, and this was prevented by the addition of glucose to the medium; pyruvate, ribose, glycerol, and L-alpha-glycerol phosphate were ineffective. An active oxidative phosphorylation system and tricarboxylic cycle appear to be present and essential in the rabbit embryo from the 1-cell stage, but glycolysis may not be essential until blastocyst formation.

Animals↗

In vitro fertilization, culture, and transfer of rabbit ova.

Ovulated rabbit oocytes were fertilized in vitro in chemically defined media supplemented with bovine serum albumin and either cultured up to the expanding blastocyst stage or transferred to recipients after varying periods of culture. Embryos transferred after up to 72 hours of in vitro culture were born as viable young. Oocytes from young virgin does were superior to oocytes from nonvirgin does for the purpose of in vitro fertilization (54% versus 26% fertilized, P less than 0.01). Capacitated sperm from artificially inseminated capacitators resulted in fertilization rates slightly lower than those from naturally mated does (46% versus 57% fertilized, P less than 0.025). Removal of cumulus and corona cells from oocytes with hyaluronidase and repeated aspiration through a fine pipette resulted in lowered fertilization rates (51% versus 73%, P less than 0.025). Linbro Disposo Tray wells were as good as glass tissue-culture dishes for the in vitro mixing of gametes and were more convenient to use. Modified Ham's F10 medium was used to culture the in vitro-fertilized embryos. However, when a modified Brackett's medium was used instead of modified Ham's F10 for the initial 4-hour period after mixing gametes, more oocytes were fertilized (52% versus 28%, P less than 0.01).

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