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337 Volume Regulation in Second Trimester Cytotrophoblast Cells
SPO Abstracts 369
Volume 166 Number I. Part 2
336 MATERNALFLOORINFARCI'ION:RELATIONSHIPOF
X-CELLS, MAJOR BASIC PROTEIN AND ADVERSE PERINATAL OUTCOME. K.K. Vernor," K. Benirschke, G.M. Kephart," T.L. Wasmoen," J.A. Ney and G,J. Gleich," Dept. ObiGyn and Immunology, Mayo Clinic, Rochester, MN and Dept. Pathology, UCSD, San Diego, CA. To expand our knowledge of pregnancy-associated major basic protein (pMBP) and the X-cell in reproductive physiology, we analyzed pregnancies complicated by maternal floor infarction (MFI). MFI placentas are grossly abnormal showing a striking increase in the number and size of subchorionic cysts and histologically show increased proliferation of X-cells. The pregnancies have poor outcomes with either intrauterine growth retardation or fetal death. Previously, pMBP has been localized to the placental X-cell and identified at elevated levels in serum and amniotic fluid in all normal pregnancies. Here, we present a classic example of MFI along with seven other cases. We analyzed placental tissue, serum, amniotic fluid, and placental cyst fluid. Serum pMBP levels were variably elevated both in normal and MFI pregnancies. Placental tissue from MFI pregnancies had increased numbers of X-cells and fibrinoid material that occupied or surrounded degenerating villi and which stained intensely for pMBP. These results indicate that pMBP, a potent cytotoxin and platelet agonist, is deposited in close proximity to chorionic villi in MFI, and may contribute to the pathophysiology of this disorder.
337
VOLUME REGUlATION IN SECOND TRIMESTER CYTOTROPHOBlAST CELLS. Robert S. Egerman,' John M. Bissonnette,' Gail B. Willeke,x Dept. Ob/Gyn, Oregon Health Sciences Univ., Portland, OR. Although ion transport systems have been described in syncytiotrophoblast brush border vesicles, no study as yet addresses cell volume regulation in placental cells. These mechanisms are important in understanding fetal H20 acquisition in physiologic and in pathologic states (e.g. hyperosmolar ketosis). Cytotrophoblast cells were isolated from 17·19 week human placentas by enzyme digestion and density gradient centrifugation. Video-microscopy captured interval cell images as cells were exposed to various solutions. These recorded images were stored in a MacIntosh computer and later recalled for area measurements using Image 1.22 software. In the continued presence of mannitol (4OOmM) shrunken cytotrophoblasts increased their area 1.8 to 3.2 fold after an 8-12 minute latency period. When extracellular NaCI was replaced with N-methyl g1ucamine, the latency period was extended and final volume was depressed. In the presence of the Na +-H + exchanger antagonist, ameloride (10Q..£ M), no increase in cell area occurred. Conclusion: 1) Cytotrophoblasts are osmotically active. 2) Na + is necessary for regulatory cell volume increase. 3) Na+ -H+ exchanger is an important system for Na + (H20) entry into the cell. 4) The latency period suggests that the Na +-H + exchanger must be activated before these cells can regain their volume.
338
THE EFFECT OF 01 FFE1lENT EXTRACELLULAR KATRIJ: PROTEINS AND DIFFERENT GERUM-FREE MEDIA ON THB OUTGROWTH TROPHODLAST CELLS ISOLATED FROM HUKAN TERM PLACENTAE
or
J _
I..cYten~
C.
DepartJCent O(
l.uYten~
R.
Obstetrics
pi jnenbor/" A. Van Asschc ,
Cynecoloqy.
University of wuven,
Hclqium In order to stu4y the interaction of trophoblast cell~ with maternal tissue, a monolayer culture systea was established. We modified the .ethods described by Kliaan et al (Endocrinoloqy 118,1561,1986) and Loke , Burland (Placenta 9,113 .. 1988). The isolated cells were grown on substrates coated with the (ollOW'ing extracellular matrix proteins (ECH-proteins) : ttatrigel (H
without
H
E 0 I A
Col.
119·
RPHI1640
+
+
HEl1-D-VAL
+
Ham's Fl2
+
Laa.
Fib.
+++
++++
++
OHEH
++
K-SfM
+++
Concerning the E01-prcteins, fibronectin, followed by lam1.nin, gives the best result. Hatrigel seems to stimulate fibroblast proliferation more than the other media do. For the serum-free media, K-SFH followed by OHEH is more favourable than Haa's f12. MEH-D-VAL and RPHI1640. Ham's F12 seems to stimulate fibroblast. proliferation more than t.he other media do.
339
SINGLE INTRAUTERINE DEMISE IN T11IN PREGNANCY.
M. Eglowstein and H. D'Alton, Department of OB/GYN, Tufts University School of Medicine, Boston, MA Eighteen twin pregnancies with one intrauterine fetal demise (!UFO) noted after the first trimester were managed using a standard protocol between January 1, 1987 and Jul y 1, 1991, during which time 367 twin del iveries occurred. Of the 17 del ivered pregnancies, pLacentation was determined in 16; 9 were diamniotic/dichorionic (DC) and 7 were diarrniotic/ monochorionic (MC). Of the DC placentations, 5 cases of !UFO were unexplained, 2 were associated with major fetal anomaL ies, 1 with nonill1T1Une hydrops and 1 with placental infarct. Of the MC placentations, IUFO in 6 cases was associated with twin-twin transfusion (TTTS); the seventh was associated with preterm rupture of merrbranes and chorioamnionitis. The interval from diagnosis of IUFO to del ;very ranged from 2 days to 11 weeks in DC placentation, and 1 day to 14 weeks in MC placentation. The management protocoL consisted of frequent ultrasound for fetal growth, weekly nons tress test and biophysical profi le after 28 weeks, and weekly coagulation profi leo There were no elective preterm del iveries. Incidence of preterm del ivery was 82.3% overall (14/17); 7 of 9 DC and all MC gestations deL ivered preterm. Estimated gestational age at del ivery ranged from 28 to 38 weeks in DC gestations and 20 to 36 weeks in MC gestations. No patient required treatment for coagulopathy. 1 case of periventricular encephalomalacia was di agnosed prenatally by ul trasound 24 hours after the death of the co-twin. This seems to indicate that the timing of this neurological insult happened before the death of the co-twin. This report confirms our previous findings of a low risk of cl inically significant maternal coagulopathy and a low incidence of periventricular encephalomalacia in co-twins of single !UFO. This series demonstrates a higher incidence of spontaneous preterm deL ivery in both types of placentation than has been previously reported.
Volume 166 Number I. Part 2
336 MATERNALFLOORINFARCI'ION:RELATIONSHIPOF
X-CELLS, MAJOR BASIC PROTEIN AND ADVERSE PERINATAL OUTCOME. K.K. Vernor," K. Benirschke, G.M. Kephart," T.L. Wasmoen," J.A. Ney and G,J. Gleich," Dept. ObiGyn and Immunology, Mayo Clinic, Rochester, MN and Dept. Pathology, UCSD, San Diego, CA. To expand our knowledge of pregnancy-associated major basic protein (pMBP) and the X-cell in reproductive physiology, we analyzed pregnancies complicated by maternal floor infarction (MFI). MFI placentas are grossly abnormal showing a striking increase in the number and size of subchorionic cysts and histologically show increased proliferation of X-cells. The pregnancies have poor outcomes with either intrauterine growth retardation or fetal death. Previously, pMBP has been localized to the placental X-cell and identified at elevated levels in serum and amniotic fluid in all normal pregnancies. Here, we present a classic example of MFI along with seven other cases. We analyzed placental tissue, serum, amniotic fluid, and placental cyst fluid. Serum pMBP levels were variably elevated both in normal and MFI pregnancies. Placental tissue from MFI pregnancies had increased numbers of X-cells and fibrinoid material that occupied or surrounded degenerating villi and which stained intensely for pMBP. These results indicate that pMBP, a potent cytotoxin and platelet agonist, is deposited in close proximity to chorionic villi in MFI, and may contribute to the pathophysiology of this disorder.
337
VOLUME REGUlATION IN SECOND TRIMESTER CYTOTROPHOBlAST CELLS. Robert S. Egerman,' John M. Bissonnette,' Gail B. Willeke,x Dept. Ob/Gyn, Oregon Health Sciences Univ., Portland, OR. Although ion transport systems have been described in syncytiotrophoblast brush border vesicles, no study as yet addresses cell volume regulation in placental cells. These mechanisms are important in understanding fetal H20 acquisition in physiologic and in pathologic states (e.g. hyperosmolar ketosis). Cytotrophoblast cells were isolated from 17·19 week human placentas by enzyme digestion and density gradient centrifugation. Video-microscopy captured interval cell images as cells were exposed to various solutions. These recorded images were stored in a MacIntosh computer and later recalled for area measurements using Image 1.22 software. In the continued presence of mannitol (4OOmM) shrunken cytotrophoblasts increased their area 1.8 to 3.2 fold after an 8-12 minute latency period. When extracellular NaCI was replaced with N-methyl g1ucamine, the latency period was extended and final volume was depressed. In the presence of the Na +-H + exchanger antagonist, ameloride (10Q..£ M), no increase in cell area occurred. Conclusion: 1) Cytotrophoblasts are osmotically active. 2) Na + is necessary for regulatory cell volume increase. 3) Na+ -H+ exchanger is an important system for Na + (H20) entry into the cell. 4) The latency period suggests that the Na +-H + exchanger must be activated before these cells can regain their volume.
338
THE EFFECT OF 01 FFE1lENT EXTRACELLULAR KATRIJ: PROTEINS AND DIFFERENT GERUM-FREE MEDIA ON THB OUTGROWTH TROPHODLAST CELLS ISOLATED FROM HUKAN TERM PLACENTAE
or
J _
I..cYten~
C.
DepartJCent O(
l.uYten~
R.
Obstetrics
pi jnenbor/" A. Van Asschc ,
Cynecoloqy.
University of wuven,
Hclqium In order to stu4y the interaction of trophoblast cell~ with maternal tissue, a monolayer culture systea was established. We modified the .ethods described by Kliaan et al (Endocrinoloqy 118,1561,1986) and Loke , Burland (Placenta 9,113 .. 1988). The isolated cells were grown on substrates coated with the (ollOW'ing extracellular matrix proteins (ECH-proteins) : ttatrigel (H
without
H
E 0 I A
Col.
119·
RPHI1640
+
+
HEl1-D-VAL
+
Ham's Fl2
+
Laa.
Fib.
+++
++++
++
OHEH
++
K-SfM
+++
Concerning the E01-prcteins, fibronectin, followed by lam1.nin, gives the best result. Hatrigel seems to stimulate fibroblast proliferation more than the other media do. For the serum-free media, K-SFH followed by OHEH is more favourable than Haa's f12. MEH-D-VAL and RPHI1640. Ham's F12 seems to stimulate fibroblast. proliferation more than t.he other media do.
339
SINGLE INTRAUTERINE DEMISE IN T11IN PREGNANCY.
M. Eglowstein and H. D'Alton, Department of OB/GYN, Tufts University School of Medicine, Boston, MA Eighteen twin pregnancies with one intrauterine fetal demise (!UFO) noted after the first trimester were managed using a standard protocol between January 1, 1987 and Jul y 1, 1991, during which time 367 twin del iveries occurred. Of the 17 del ivered pregnancies, pLacentation was determined in 16; 9 were diamniotic/dichorionic (DC) and 7 were diarrniotic/ monochorionic (MC). Of the DC placentations, 5 cases of !UFO were unexplained, 2 were associated with major fetal anomaL ies, 1 with nonill1T1Une hydrops and 1 with placental infarct. Of the MC placentations, IUFO in 6 cases was associated with twin-twin transfusion (TTTS); the seventh was associated with preterm rupture of merrbranes and chorioamnionitis. The interval from diagnosis of IUFO to del ;very ranged from 2 days to 11 weeks in DC placentation, and 1 day to 14 weeks in MC placentation. The management protocoL consisted of frequent ultrasound for fetal growth, weekly nons tress test and biophysical profi le after 28 weeks, and weekly coagulation profi leo There were no elective preterm del iveries. Incidence of preterm del ivery was 82.3% overall (14/17); 7 of 9 DC and all MC gestations deL ivered preterm. Estimated gestational age at del ivery ranged from 28 to 38 weeks in DC gestations and 20 to 36 weeks in MC gestations. No patient required treatment for coagulopathy. 1 case of periventricular encephalomalacia was di agnosed prenatally by ul trasound 24 hours after the death of the co-twin. This seems to indicate that the timing of this neurological insult happened before the death of the co-twin. This report confirms our previous findings of a low risk of cl inically significant maternal coagulopathy and a low incidence of periventricular encephalomalacia in co-twins of single !UFO. This series demonstrates a higher incidence of spontaneous preterm deL ivery in both types of placentation than has been previously reported.