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PURPOSE: The aim of this study was to establish an in vitro method to purify human multipotent adult progenitor cells (hMAPCs) and assess their possible differentiation into hepatocytes by coculture with human hepatocyte line L02. METHODS: hMAPCs were isolated by magnetic activated cell sorting (MACS) depletion selection using CD45 and GlyA microbeads. After indirect or direct coculture of hMAPCs and human hepatocyte line L02, the expression of albumin (ALB), alpha-fetoprotein (AFP), cytokeratin (CK) 18, and CK19 by hMAPCs was detected by immunocytochemistry. RESULTS: With the MACS method, (5-10) x 10(4)/mL hMAPCs could be separated from 1 x 10(6)/mL bone marrow mononuclear cells. The purity of CD45-/GlyA- cells separated from bone marrow adherent cells was more than 98%, as determined by flow cytometry. In the coculture without cell-to-cell contact, hMAPCs expressed high AFP on day 1, and then tapered daily to low expression on day 7; ALB expression reached its peak on day 5, and remained high on day 7; CK18 was initially expressed on day 5 and was higher on day 7; CK19 was negative in all assays. In the coculture with cell-to-cell contact, ALB and CK18 were expressed by most cells while AFP appeared in only a few on day 5. CONCLUSION: hMAPCs were induced to differentiate into mature hepatocyte-like cells by coculture with a hepatocyte cell line, either with or without cell-to-cell contact, but the former seemed more effective.
Subject(s)
Adult , Humans , alpha-Fetoproteins , Bone Marrow , Cell Differentiation , Cell Line , Coculture Techniques , Flow Cytometry , Hepatocytes , Immunohistochemistry , Keratins , Microspheres , Stem CellsABSTRACT
Objective To find a better composition substitution fluid for continuous venovenous hemofiltration (CWH) comparing with the Port's composition substitution fluid,and to evaluate its efficiency and prognosis on patients with dysfunction of glycometabolism and multiple organ dysfunction syndrome (MODS) caused by trauma. Method Totol of 134 patients in the Firist Affiliated Hospotal of the Fourth Miliatary Medicial University,with glycometabolism dysfunction and MODS caused by trauma between Janurary 2001 and December 2006 were treated with CVVH.They were divided into two groups:the Port's composition substitution fluid which glucose concentration was 59.1 mmoL/L (group A,59 cases) and the new substitution fluid which glucose concentration is 11.8 mmol/L (group B,75 cases).The changes of electrolytes,arterial blood gas analysis,blood glucose and osmolarity during CVVH were observed.Results The mortality in group A was significantly higher than that in group B (64.4 % vs 40.0 %,P = 0.005).Before treated with CVVH,there was no significant differences of APACHEⅡscores between group A and group B.Either in the survival patients or in non-survivors,the APACHEⅡscores in group B were significantly higher than that in group A (P
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1 ml/kg)and the ICU stay.Results The hospital mortality of AKI phaseⅢwas significantly higher than that of AKI phaseⅠorⅡ(P
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Objective To propose a new blood purification modality-hemodialysis with plasma- based dialysate (HD-PBD) plus high volume hemofiltration (HVHF) for patients with liver failure, and to evaluate the effect of this treatment on plasma cytokines.Methods Twelve patients with liver failure were included in this study.All patients received HD-PBD therapy in the first 6 hours,and then were treated with HVHF for 24 hours with the same filter (AV600).The levels of TNF-?,IL-1?, IL-6 and IL-8 in plasma before and after HD-PBD plus HVHF for 6 and 24 hours were examined respectively by ELISA,and changes of clinical parameters were observed at the same time point. Serum bilirubin,total bile acids (TBA),serum ammonia,blood urea nitrogen (BUN) and serum creatinine (Scr) were detected before and after treatment.Arterial blood gas analysis and the concentration of electrolytes were monitored before and after treatment.Results (1)HD-PBD for 6 hours was more effective than HVHF for 24 hours in removal of serum bilirubin and TBA(P＜0.05). (2)Serum ammonia,BUN,Ser,arterial blood HCO_3~-,PCO_2,PO_2 and electrolytes did not show significant difference before and after HD-PBD (P＞0.05),but these parameters significantly changed before and after HVHF (P＜0.05).(3)The average level of serum bilirubin was sharply decreased after HVHF for 24 h following HD-PBD(P＜0.05).(4)After HD-PBD plus HVHF,there was a marked reduction of the plasma levels of TNF-?,IL-6 and IL-8.Conclusions HD-PBD plus HVHF,a newly proposed modality for patients with liver failure,can effectively decrease serum bilirubin,TBA,BUN,Scr,ammonia and cytokines,and adjust water-electrolyte as well as acid- alkali balance.It is a low-cost,safe,simple and convenient therapy.