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Cobe 2991 cell processor

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The COBE 2991 Cell Processor is a lab equipment product designed for processing biological samples. It is capable of centrifugation and separation of cellular components from biological fluids. The device's core function is to isolate and concentrate specific cell types or components for further analysis or processing.

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Lab products found in correlation

Human serum albumin (hsa), by Baxter (2 mentions) Insulin like growth factor 1, by Cell Sciences (2 mentions) Biocoll, by Harvard Bioscience (2 mentions) Ham s f10 based medium, by Lonza (2 mentions) Eclipse ti microscope, by Nikon (1 mentions) Nucleocounter yc 100, by ChemoMetec (1 mentions) Neutral protease, by Serva Electrophoresis (1 mentions) Collagenase, by Serva Electrophoresis (1 mentions)

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COBE 2991 (6 citations)

6 protocols using cobe 2991 cell processor

1

Islet Isolation and Culture Protocol

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The cannulated pancreas was perfused with enzyme solution using an automatic perfusion apparatus (BioRep Technologies, Miami, FL, USA). The pancreas was then cut into 6–9 pieces and placed into the Ricordi chamber for digestion in the presence of 7 marbles. Tissue digestion was conducted at 37°C using a previously described method.21 The digested pancreatic tissue was collected for purification of the islets using a refrigerated COBE 2991 Cell Processor (Terumo BCT, Inc., Lakewood, CO, USA). The islets were washed and prepared for culture immediately after the isolation process. Islets were cultured in Connaught Medical Research Laboratories (CMRL)-1066 Supplemented media (pH 7.4) with 0.5% human serum albumin (Baxter Healthcare Corporation, Irvine, CA, USA) and 0.1 μg/mL insulin-like growth factor-1 (Cell Sciences, Canton, MA, USA).
Prior to placing the islets in culture, islets were re-suspended in 100mL of CMRL culture media and a second sterility sample (post-isolation) was obtained for testing. Islets were cultured at 22°C/5%CO2 for 24–72 hrs before the final islet product was collected for Quality Control Assessment.22 (link) At this point, a third sample (post-culture) was taken for sterility testing. The entire isolation process, islet culture, and collection of the final product were carried out in the cGMP facility.
Qi M., Omori K., Mullen Y., McFadden B., Valiente L., Juan J., Bilbao S., Tegtmeier B.R., Dafoe D., Kandeel F, & Al-Abdullah I.H. (2016). Prophylactically Decontaminating Human Islet Product for Safe Clinical Application: Effective and Potent Method. Transplantation Direct, 2(2), e63.
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2

Islet Isolation and Culture Protocol

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The cannulated pancreas was perfused with enzyme solution using an automatic perfusion apparatus (BioRep Technologies, Miami, FL). The pancreas was then cut into 6 to 9 pieces and placed into the Ricordi chamber for digestion in the presence of 7 marbles. Tissue digestion was conducted at 37°C using a previously described method.21 The digested pancreatic tissue was collected for purification of the islets using a refrigerated COBE 2991 Cell Processor (Terumo BCT, Inc., Lakewood, CO). The islets were washed and prepared for culture immediately after the isolation process. Islets were cultured in Connaught Medical Research Laboratories 1066 Supplemented media (pH 7.4) with 0.5% human serum albumin (Baxter Healthcare Corporation, Irvine, CA) and 0.1 μg/mL insulin-like growth factor-1 (Cell Sciences, Canton, MA).
Before placing the islets in culture, islets were resuspended in 100 mL of Connaught Medical Research Laboratories culture media, and a second sterility sample (postisolation) was obtained for testing. Islets were cultured at 22°C/5% CO2 for 24 to 72 hours before the final islet product was collected for quality control assessment.22 (link) At this point, a third sample (postculture) was taken for sterility testing. The entire isolation process, islet culture, and collection of the final product were carried out in the cGMP facility.
Qi M., Omori K., Mullen Y., McFadden B., Valiente L., Juan J., Bilbao S., Tegtmeier B.R., Dafoe D., Kandeel F, & Al-Abdullah I.H. (2016). Prophylactically Decontaminating Human Islet Product for Safe Clinical Application: Effective and Potent Method. Transplantation Direct, 2(2), e63.
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3

Islet Cell Isolation and Characterization

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Islet cells were isolated using a modification of the automated Ricordi method17
and purified by continuous gradient with Biocoll (Biochrom, Berlin, Germany) and a cooled COBE 2991 cell processor (Terumo BCT, Lakewood, CO, USA). After isolation and purification, the cell preparations were cultured in cell culture flasks T175 (Sarstedt, Nümbrecht, Germany) at 37°C in a humidified incubator (5% CO2) in a Ham’s F10 based medium (Lonza, Bazel, Switzerland).
Preparations were characterized immediately after purification and after a 1- to 5-day culture period by their beta cell number, insulin content, and insulin purity, as described previously17
. Beta cell number was calculated from the total nuclear count (NucleoCounter YC-100; ChemoMetec, Allerod, Denmark) and the percentage of insulin positive cells [immunocytochemistry with guinea pig anti-insulin (1/2,000, in-house produced) on 1.5 µm araldite sections; >2 × 103 cells counted; pictures were captured using Nikon Eclipse Ti microscope and analyzed with NIS-Elements AR v5.21 software (Nikon Europe, Amsterdam, The Netherlands; Fig. 1)]. Yield immediately post purification was also expressed as islet equivalent (IEQ), calculated using a volume based method after dithizone staining19
.
De Paep D.L., Van Hulle F., Ling Z., Vanhoeij M., Hilbrands R., Distelmans W., Gillard P., Keymeulen B., Pipeleers D, & Jacobs-Tulleneers-Thevissen D. (2022). Utility of Islet Cell Preparations From Donor Pancreases After Euthanasia. Cell Transplantation, 31, 09636897221096160.
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4

Islet Isolation and Transplantation Protocol

Cited 2 times
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Islets were isolated by collagenase digestion with VitaCyte intact C1 collagenase (VitaCyte, Indianapolis, IN) and SERVA neutral protease (SERVA, Heidelberg, Germany), followed by mechanical digestion using the semi-automated method of Ricordi, as previously described (16 (link), 17 (link)). Purification with density centrifugation using a COBE 2991 cell processor (Terumo BCT, Lakewood, CO) was performed only if needed to reduce tissue volume in cases where volume initially exceeded 0.25 mL per kilogram body weight. This was the case in three out of 26 patients in this series. Islets were suspended in one or more bags of 200 mL CMRL culture medium (Mediatech, Inc., Manassas, VA). Islets were prepared with heparin in the islet product and heparin was also administered to the patient (total dose 70 units/kg body weight) to reduce risk for portal vein thrombosis. Islets were infused by gravity into the portal vein. Portal pressures were monitored during infusion, and in cases where portal pressures exceed 25 cm saline, the remainder of the islet product was implanted in the peritoneal cavity as a thin film (6 (link)).
McEachron K.R., Skube M.E., Yang Y., Hodges J.S., Wilhelm J., Beilman G., Chinnakotla S., Schwarzenberg S.J, & Bellin M.D. (2019). Utility of arginine stimulation testing in preoperative assessment of children undergoing total pancreatectomy with islet autotransplantation. Clinical transplantation, 33(8), e13647.
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5

Islet Cell Isolation and Culture Protocol

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Islet cells were isolated using a modification of the automated Ricordi method [22 (link)]. During the study period, three types of enzyme were used for organ distention and digestion but only organs processed using NB1 collagenase (Serva Electrophoresis, Heidelberg, Germany) were included for analysis to exclude an effect of enzyme type on isolation outcome. Different batches of NB1 collagenase were evenly distributed between donor types, minimizing an effect of batch-to-batch variability on outcome. Isolates were purified by continuous gradient with Biocoll (Biochrom, Berlin, Germany) and a cooled COBE 2991 cell processor (Terumo BCT, Lakewood, CO, USA). After isolation and purification, the cell preparations were cultured in TC suspension culture flasks T175 (Sarstedt, Nümbrecht, Germany) at 37°C in a humidified incubator (5% CO2) in a Ham’s F10 based medium (Lonza, Bazel, Switzerland) or CMRL based medium (Mediatech, Manassas, VA, USA) [22 (link)].
De Paep D.L., Van Hulle F., Ling Z., Vanhoeij M., Pirenne J., Keymeulen B., Pipeleers D, & Jacobs-Tulleneers-Thevissen D. (2021). Lower beta cell yield from donor pancreases after controlled circulatory death prevented by shortening acirculatory warm ischemia time and by using IGL-1 cold preservation solution. PLoS ONE, 16(5), e0251055.
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6

Isolation of Human Pancreatic Islets

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Human islets were isolated from cadaver donors following a protocol based on that described in Ricordi et al. (20) . Briefly, a mixture of a collagenase and neutral protease (Serva, Heidelberg, Germany) was instilled into the main duct of the pancreas. collagenase from either of the two producers was used (VitaCyte, Indianapolis, IN, USA). After incubation in a Ricordi chamber (20), the islets were separated from the exocrine tissue via centrifugation using a COBE 2991 Cell Processor (Terumo BCT, Tokyo, Japan) and continuous Ficoll gradient (Biochrom, Cambridge, UK). The isolated islets were stabilized in a CMRL-based medium. Routine test samples were taken to assess graft quantity and quality.
Habart D., Švihlik J., Schier J., Cahová M., Girman P., Zacharovová K., Berková Z., Kříž J., Fábryová E., Kosinová L., Papáčková Z., Kybic J, & Saudek F. (2016). Automated Analysis of Microscopic Images of Isolated Pancreatic Islets. Cell transplantation, 25(12).
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