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Identification of trophoblastic cells by means of a fluorochrome method
Identification of trophoblastic cells by means of a fluorochrome method R.
PIMENTA
M.
R.
Q.
HILTON Rio
DE DE VIANA,
de Janiero,
MELLO,
KASTNER,
M.D PH.D.
M.D.
Brazil
T H E identification of syncytiotrophoblast in the umbilical cord and peripheral blood of pregnant women is not a simple task. Several articles in the medical literature refer to the presence of syncytiotrophoblastic cells, but the results have been contradictory. 1, 2, 8, 4 The purpose of this paper is to report additional observations and to evaluate the fluorescence microscopic technique. Material
and
were taken from the antecubital vein. The previously published fluorescence method’ for peripheral blood studies was followed. Others’ slides were used for the study with hemalum and eosin-azure blue and hematoxylin and eosin. Results
Placental imprints were revealed with fluorochrome dye acridine-orange (AO) typical trophoblastic cells. The striking flaming red fluorescence is based on the fact that those cells contain a larger amount of cytoplasmic ribonucleic acid, while the DNA of the nucleus appears in the form of yellowgreen fluorescence. Since acridine orange is a dye which quantitatively demonstrates the amount of RNA and DNA in all cells, the cells of the syncytiotrophoblast cannot be mistaken for the megakaryocytes.8
methods
The material for this study was comprised of 46 patients at various stages of gestation (Table I). Examinations were first carried out on samples from the placental site immediately following delivery. The freshly cut thin section is touched lightly to clean slides and fixed. The smears are stained according to the fluorescence microscopic technique developed by von Bertalanffy, Masin, and Masin based on differential analysis of ribonucleic acid (RNA) and desoxyribose nucleic acid (DNA) . This specific histochemical reaction has been already tested in tumor imprints.6 Duplicate smears were processed according to the hematoxylin and eosin technique or hemalum and eosin-azure blue method. From the 46 patients blood specimens
Table I. Stage of gestation
of patients
in
the study Pregnancy
(wk.)
No.
12 16 20 24 28 32
From the Laboratory of Experimental Pathology, Hospital Evandro Chagas Instituto Oswald0 Crur.
Total
606
case*
of
a
1 5 14 10 7 3 6 46
-
Volume
89
?;umber
5
Trophoblastic
cell
identification
607
The slides of peripheral blood were screened by the authors to demonstrate the presence of trophoblastic cells, such as those seen in imprints of the placental site. There was not a single trophoblastic cell found. Summary
Fig. 1. Cytoplasmic trophoblast has been dye acridine orange. present.
1.
2.
3.
basophilia in the placental shown by the fluorochrome Several lymphocytes are also
1. The fluorescence method can be used for the rapid screening for cells of the syncytiotrophoblast. 2. The syncytiotrophoblast contains a large amount of RNA in the cytoplasm. 3. Syncytiotrophoblasts were demonstrated in placental imprints, but they were not detected in the blood from the peripheral circulation.
REFERENCES
4.
Douglas, N. M.,
5.
G. W., Thomas, L., Carr, M., Cullen, and Morris, R.: AM. J. OBST. & GYNEC. 78: 960, 1959. Hertin, A. T., Adams, E. C.. McKay. D. G., Rock;j., Mulligan, W: J., a& Menk&, M. F.I AM. T. OBST. & GYNEC. 75: 1025. 1958. Thie& H. A.: AM. J. OBST. & ~YNEC. 79: 636, 1960.
6. 7. 8.
Salvaggio, A. T., Nigogosyam, G., and Mack, H. C.: AM. J. OBST. & GYNEC. 80: 1013, 1960. von Bertalanffy, L., Masin, M., and Masin, F. A.: Cancer 11: 873, 1958. Pimenta de Mello, R., Kastner, M. R. Q., and Tavares, B. M.: 0 Hospital 62: 747, 1962. Pimenta de Mello, R.: Acta cytol. 7: 62, 1963. Pimenta de Mello, R., Santos, L. G., and Kastner, M. R. Q.: Acta cytol. 7: 66, 1963.
PIMENTA
M.
R.
Q.
HILTON Rio
DE DE VIANA,
de Janiero,
MELLO,
KASTNER,
M.D PH.D.
M.D.
Brazil
T H E identification of syncytiotrophoblast in the umbilical cord and peripheral blood of pregnant women is not a simple task. Several articles in the medical literature refer to the presence of syncytiotrophoblastic cells, but the results have been contradictory. 1, 2, 8, 4 The purpose of this paper is to report additional observations and to evaluate the fluorescence microscopic technique. Material
and
were taken from the antecubital vein. The previously published fluorescence method’ for peripheral blood studies was followed. Others’ slides were used for the study with hemalum and eosin-azure blue and hematoxylin and eosin. Results
Placental imprints were revealed with fluorochrome dye acridine-orange (AO) typical trophoblastic cells. The striking flaming red fluorescence is based on the fact that those cells contain a larger amount of cytoplasmic ribonucleic acid, while the DNA of the nucleus appears in the form of yellowgreen fluorescence. Since acridine orange is a dye which quantitatively demonstrates the amount of RNA and DNA in all cells, the cells of the syncytiotrophoblast cannot be mistaken for the megakaryocytes.8
methods
The material for this study was comprised of 46 patients at various stages of gestation (Table I). Examinations were first carried out on samples from the placental site immediately following delivery. The freshly cut thin section is touched lightly to clean slides and fixed. The smears are stained according to the fluorescence microscopic technique developed by von Bertalanffy, Masin, and Masin based on differential analysis of ribonucleic acid (RNA) and desoxyribose nucleic acid (DNA) . This specific histochemical reaction has been already tested in tumor imprints.6 Duplicate smears were processed according to the hematoxylin and eosin technique or hemalum and eosin-azure blue method. From the 46 patients blood specimens
Table I. Stage of gestation
of patients
in
the study Pregnancy
(wk.)
No.
12 16 20 24 28 32
From the Laboratory of Experimental Pathology, Hospital Evandro Chagas Instituto Oswald0 Crur.
Total
606
case*
of
a
1 5 14 10 7 3 6 46
-
Volume
89
?;umber
5
Trophoblastic
cell
identification
607
The slides of peripheral blood were screened by the authors to demonstrate the presence of trophoblastic cells, such as those seen in imprints of the placental site. There was not a single trophoblastic cell found. Summary
Fig. 1. Cytoplasmic trophoblast has been dye acridine orange. present.
1.
2.
3.
basophilia in the placental shown by the fluorochrome Several lymphocytes are also
1. The fluorescence method can be used for the rapid screening for cells of the syncytiotrophoblast. 2. The syncytiotrophoblast contains a large amount of RNA in the cytoplasm. 3. Syncytiotrophoblasts were demonstrated in placental imprints, but they were not detected in the blood from the peripheral circulation.
REFERENCES
4.
Douglas, N. M.,
5.
G. W., Thomas, L., Carr, M., Cullen, and Morris, R.: AM. J. OBST. & GYNEC. 78: 960, 1959. Hertin, A. T., Adams, E. C.. McKay. D. G., Rock;j., Mulligan, W: J., a& Menk&, M. F.I AM. T. OBST. & GYNEC. 75: 1025. 1958. Thie& H. A.: AM. J. OBST. & ~YNEC. 79: 636, 1960.
6. 7. 8.
Salvaggio, A. T., Nigogosyam, G., and Mack, H. C.: AM. J. OBST. & GYNEC. 80: 1013, 1960. von Bertalanffy, L., Masin, M., and Masin, F. A.: Cancer 11: 873, 1958. Pimenta de Mello, R., Kastner, M. R. Q., and Tavares, B. M.: 0 Hospital 62: 747, 1962. Pimenta de Mello, R.: Acta cytol. 7: 62, 1963. Pimenta de Mello, R., Santos, L. G., and Kastner, M. R. Q.: Acta cytol. 7: 66, 1963.