ABSTRACT
【Objective】 To investigate the activity level of coagulation factor Ⅺ(FⅪ) in IVIG products, which would provide support for improving the safety and quality control of IVIG products in China. 【Methods】 A total of 71 batches of IVIG products(half of which were closed to expiry date) from 23 domestic manufacturers (brands) were submitted for inspection and assayed for procoagulant activity including FⅪa, FⅪ and NAPTT activity with a chromogenic assay, coagulation proenzymes levels using one stage clotting assays and non-activated partial thromboplastin time (NAPTT). 【Results】 FⅪa(mIU/mL): 32 lots of IVIGs from 15 manufacturers(brands) possessed activities below 0.1 or the detection limit of the assays, 24 IVIG lots from 12 manufacturers were between 0.1~1(0.39±0.20), the remaining 15 IVIG lots from 6 manufacturers had FⅪa activity greater than 1 up to 47(13.27±15.61). FⅪ(mIU/mL): there were no detectable activities of FⅪ in 58 lots from 21 manufacturers, the other 13 lots from 6 manufacturers had IVIGs between 13~309(69.0±98.1), which included in 15 lots of IVIG whose FⅪa activity were greater than 1mIU/mL. NAPTT(s): The coagulation times for aforementioned 15 IVIG lots were all above 150s in the NAPTT test. The NAPTT ratios varied between 0.67 and 0.94(0.83±0.07), and 7 of 15 IVIGs had NAPTT ratios below or equal to 0.8. 【Conclusion】 There are substantial differences in the FⅪ and FⅪa activities in the IVIG preparations from 23 different manufacturers. Most of them had a lower activities; certain lots from specific manufacturers had relatively higher FⅪa activities. Manufactures should evaluate the manufacturing processes and monitor IVIG preparations for the presence of factor Ⅺ-like activity in case of thrombotic risks.
ABSTRACT
Objective To investigate the effects of different culture conditions on the isolation and expansion of stem cells from second-trimester amniotic fluids.Methods Amniotic fluids were obtained from 15 pregnant women undergone amniocenteses for medical indications between 16-24 gestation weeks by transabdominal amniocenteses from September 2007 to June 2008.Amniotic fluids(10-20 ml)samples were collected and each WaS cultured under different conditions or groups.(1)Low-glucose DMEM(LD) medium supplemented with 10%of fetal bovine serum(group of 10% FBS);(2)LD medium with 20%of FBS(group of 20%FBS);(3)LD medium with 15%of FBS and 4 ng/ml of basic fibroblast growth factor (group of bFGF);(4)LD medium with 10%of FBS as well ag the culture plate coated with gelatin(group of gelatin).The effects of different conditions were evaluated by comparing the number of primary colonies,the cell morphology and the ability of expansion.The isolated stem cells were identified by flow cytometry,RT-PCR and differentiation ability to edipocyte.Resuits (1)The success rates of primary culture of the group of 10%FBS,20%FBS,bFGF and gelatin were 60%,73%,73%and 60% respectively(P>0.05).The numbers of colonies were 0.9±0.5,2.6±1.5,2.9±1.5,1.1±0.8(P<0.01 when group of 10%FBS and gelatin compared with group of 20%FBS and bFGF);among the primary colonies,fibroblast-like colonies accounted for 46%,49%,64%.44%respectively(P>0.05).(2)The second passage cells obtained from all of these four groups could difierentiate into adipocyte after induction.(3)In the group of bFGF,stem cells were isolated from 5 samples and expanded to nearly 107 cells after 5 passages(P<0.01 compared with other groups).(4)Karyotype were normal in all samples.(5)Stem cells from bFGF group showed positive expression of SSEA-4.Oct-4 and Nanog gene detected by flow cytometry and RT-PCR.Conclusion Stem cells can be isolated from second-trimester amniotic fluids;moderate serum concentration and supplementation of bFGF can improve the efficiency of isolation and expansion of amniotic fluid of stem cells.