ABMSC Infusion Through Hepatic Artery in Portal Hypertension Surgery for the Treatment of Liver Cirrhosis
ABMSC Infusion Through Hepatic Artery in Portal Hypertension Surgery for the Treatment of Liver Cirrhosis
In recent years, the safety and efficacy of autologous bone marrow stem cells infusion
 (ABMSCi) therapy were confirmed. The investigators attempted to infuse autologous bone marrow
 stem cells (ABMSC) through inserting a catheter into right gastric artery as far as proper
 hepatic artery after finishing open abdominal portal hypertension surgery. The present study
 was designed to treat bleeding from esophageal varices and hypersplenism and hopefully to
 improve the liver function as well.
1. ABMSC mobilization and harvest

 -  For harvesting more ABMSC, ABMSC mobilization was induced by rhG-CSF (Gran○R),
 administered subcutaneously at a dose of 300μg daily for three consecutive days
 before open abdominal portal hypertension surgery.

 -  Bone marrow (160-200ml) of the patients was harvested from both posterior superior
 iliac according to standard procedures under local anaesthesia and was collected in
 a plastic bag containing heparin.

 2. Open abdominal portal hypertension surgery

 - Immediately after the harvest of ABMSC, the modified Sugiura procedure was performed
 for the patients who were assigned to the study group. The same surgical procedure was
 also performed for the control group (without harvest of ABMSC and ABMSC infusion).

 3. ABMSC separation and infusion

 -  While performing the portal hypertension surgery, ABMSC was separated and purified
 in a class 10,000 clean laboratory. After fat and bony particles were removed by
 filtration, collected cells were moved to a cell-processing device. We used the
 reagent kit ([Patent Number] ZL 2006 1 0106875.5; [Number of Criteria Applicable]
 YZB/NING YIN 0008-2008; [Researcher and Developer] Wealthlin Science & Technology
 Inc., Canada; [Producer] Ningxia Zhonglianda Biotech Co., Ltd.). The reagents adopt
 the method of negative cells collection. Take the cells which intended to remove as
 target cells, and carry out the removal step-by-step. On the basis of this method,
 red blood cells, blood platelets, blood plasma will be completely removed with part
 of white cells and lymphocytes being remarkably removed as well while all the stem
 cells / progenitor cells are being well retained.

 -  The nucleated cell (white blood cell) count of final ABMSC was measured by an
 automated complete blood count instrument and flow cytometry analysis. The number
 of mononuclear cells was counted manually under a microscope by Wright-Giemsa stain
 method. CD34 positive cells were determined by flow cytometry analysis.

 -  The time of ABMSC separation and purification was 2.5-3 hours which had to be
 completed before the conclusion of portal hypertension surgery (3-3.5 hours). ABMSC
 was added to 10 ml saline and well mixed by shaking the vial gently. Before
 incision closure, the right gastric artery or right gastroepiploic artery was
 selected and a catheter was inserted. The catheter was pushed to reach the proper
 hepatic artery. The diameter of the catheter is 1.4mm, it is thin enough to easily
 been inserted to right gastric artery or right gastroepiploic artery (central
 venous catheterization, REF product NO.ES-04218, Arrow International, Inc.). The
 mixture of saline and AMBC was infused into hepatic artery at uniform speed for
 about two minutes. The catheter was removed after the ABMSCi. The puncture point of
 the right gastric artery was repaired using blood vessel suture or transfixed.

 4. Statistical analysis - Categorical data are presented as absolute values and
 percentages, whereas continuous data are summarized as mean and Standard Deviation.
 Statistical analysis was performed using t-test for paired or unpaired samples. Time
 courses of measurements of liver function parameters were analyzed by repeated-measures
 ANOVA. The analyses were performed using the SPSS 15.0 statistical package (SPSS Inc.,
 Chicago, IL, USA). All statistical analyses were based on two-tailed hypothesis tests
 with a significance level of p< 0.05.
Inclusion Criteria:

 1. Advanced liver cirrhosis after hepatitis B resulted in bleeding from esophageal
 varices and hypersplenism, and needed open abdominal portal hypertension surgery;

 2. Endoscopy evidence of showing severe gastric and esophageal varices;

 3. Severe hypersplenism （white blood cells (WBC) <3×109/L and platelet (PLT) <100×109/L）;

 4. Active bone marrow hyperplasia showed by bone marrow biopsy before surgery;

 5. Age between 18 and 60 years;

 6. Plasma albumin <35g/L, or mild ascites;

 Exclusion Criteria:

 1. Enlisted for liver transplantation

 2. Diagnosis of hepatocellular carcinoma or other cancers

 3. Other severe medical disease, and acute infection.