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PERFUSION OF HUMAN PLACENTA AS A MODEL OF VASCULAR RESEARCH

Pregnancy is the unique, short reproductive event with far reaching

consequences for the individual, family and society. The placenta is the

primary fetal organ of respiration, metabolism, nutrition and

excretion. The early detection of placental dysfunction is critical for

the fetal and maternal survival. Additionally the placenta is a highly

vascular organ and has been used as a model for brain research and

vascular responses.

The structure and function of the placenta differs between male and

female fetuses, which makes it a perfect model to study gender-

dependent vascular responses. There are indeed obvious gender

differences in vascular responsiveness and vascular metabolism, which

has been identified in other vascular beds. The technique of placental

perfusion in vitro provides a unique opportunity to evaluate the

physiological responses to the vascular active substances and estimate

the feto-maternal drug transfer.

Abstract

Maira Carrillo1, James Maher1, Gary Ventolini1, Ailena Mulkey2, Moss Hampton1

and Natalia Schlabritz-Loutsevitch1*

1Dept. of Obstetrics and Gynecology, Texas Tech University Health Sciences Center, Odessa

2Clinical Research Institute, Odessa, TX

*Natalia.schlabritz-lutsevitch@ttuhsc.edu

The objective is to establish the model of perfusion of human placenta

in-vitro and to record indirectly placental vessel contractility.

Objectives

After obtaining the informed consent, placenta was received after non-

complicated cesarean section delivery, the fetal artery was

immediately cannulated, followed by the cannulation of fetal vein

(Figure1 A). The cannulation and flushing re-established the circulation

in one of the intact cotyledons (Figure 1 B) at which time it is placed

into the perfusion chamber (Figure 1 B). The re-established circulation

is open circle fetal circulation (Figure 2). A basal fetal arterial

hydrostatic pressure (FAHP) was recorded (Figure 3). After baseline is

established, 3M KCl is added to induce vasoconstriction pressure is

recorded using Lab Chart software (ADInstruments,U.S, Colorado

Springs, Colorado). In addition to recording FAHP we also measure and

record: temperature, pH, oxygen tension using Lab Chart. We also used

the FLIR1 infrared technology to examine temperature of the sample

which also provides indication that the placenta is still metabolically

active.

Materials and Methods

The mean FAHP was 35.3±5 mmHg, mean weight of the cotyledons was

44.0±4 g. The temperature in the perfused cotyledon was constant

throughout the experiment.

Results

Conclusions

• We were able to establish placental perfusion in vitro in our

laboratory, the only placental perfusion lab in West Texas.

• Our data is in line with those published by Dr. Brownbill.

• We were able to establish a baseline FAHP and subsequently induce

vasoconstriction using 3M KCl. Induction of vasoconstriction shows

that the placenta is viable and metabolically active.

• Placenta perfusion is an excellent tool for the study fetal gender

specific responses.

Acknowledgements

• This study was supported by the funds of the Department of

Obstetrics and Gynecology TTUHSC at the PB and regional Dean

start-up funds to NSL.

• We acknowledge help of the Clinical Research institute in IRB

preparation, patients’ management and consents.

• The critical advise and help of Dr. Paul Brownbill (University of

Manchester, UK) is highly appreciated.

• We are deeply thankful medical staff and patients of MCH for their

incredible support and understanding of this study.

Patient Gestational Age (weeks) Race

Maternal age (yrs ) BMI feta l sex

Feta l wt. (g)

Cotyledon Wt. (g)

Feta l Pressure (mmHg)

1 39 Hispanic 25 28.21 female 3900 68.73 6.64

2 39 Hispanic 24 30.01 female 2830 30.4 19.46

3 29 Hispanic 34 25.31 male 3580 45.61 36.28

4 39 Hispanic 25 31.88 female 3330 63.2 38.67

39 Black 23 24.20 female 2960 27.96 51.35

1 39 Hispanic 32 36.29 male 3480 43.2 33.23

2 39 Hispanic 24 25.76 male 3680 36.94 42.62

3 39 Hispanic 27 28.54 male 3330 35.6 53.78

4 39 White 23 36.15 male 3740 45.34 -

Figure 2. Perfusion system. Orange arrows show fetal and maternal reservoirs. Blue arrows show

fetal and maternal peristaltic pumps. Green arrow heated water bath. Turquoise arrow perfusion

chamber. Pink arrow syringe pump. Yellow arrow: tank for gassing maternal and fetal perfusate.

Figure 3. Pressure recording of FAHP at baseline measurement and induction of

vasoconstriction using 3M KCl.

0

10

20

30

40

50

60

20 30 40 50 60 70 80

Basa

l fe

tal art

eri

al

hydro

stati

c

pre

ssure

(m

mH

g)

Perfused cotyledon wet weight (g)

Figure 4. Graphs showing Cotyledon weight vs FAHP ( Fetal arterial hydrostatic pressure)

A) Data collected from our laboratory.B) Data collected from Laboratory of Dr. Brownbill (University of

Manchester, UK)

Table 1. Patient, fetal and placental information.

Patient

Gestational

age

(weeks)

Maternal

age (yrs) BMI fetal sex

Newborn’s weight

(g)

Cotyledon

Wt. (g)

Fetal

Pressure

(mmHg)

1 39 25 28.21 female 3900 68.73 6.64

2 39 24 30.01 female 2830 30.4 19.46

3 29 34 25.31 male 3580 45.61 36.28

4 39 25 31.88 female 3330 63.2 38.67

5 39 23 24.20 female 2960 27.96 51.35

6 39 32 36.29 male 3480 43.2 33.23

7 39 24 25.76 male 3680 36.94 42.62

8 39 27 28.54 male 3330 35.6 53.78

9 39 23 36.15 male 3740 45.34 -

Mean±SEM 38 26 29.60±1 3386.25±112 43.96±13 35.25±5

KCl

Figure 1. Cannulation of vessels and corresponding cotyledon. A and C) Chorionic plate (A) where

vessels are cannulated (C) (blue and orange arrow) B and D) Decidual surface (B) (maternal side) after

isolation of placental lobule and cotyledon perfusion (D). Orange arrows point to temperature sensor.

Turquiose arrow shows oxygen sensor E and F) temperature map of the maternal side of placenta in the

perfusion chamber just prior to (E) and after (F) perfusion. G) Doppler image of perfused placental

cotyledon in vitro, yellow arrow shows perfusate flowing through a vessel.

Time (seconds)

Baseline

A C

B D

E

F

G

B A