PMC7563026_01
(A) Baseline continuous glucose monitoring. (B) Day 75 continuous glucose monitoring.
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There was no time spent in hypoglycemia comparing to 2% before islet graft (Fig. 1B).
PMC7563026_02
(A) Baseline continuous glucose monitoring. (B) Day 75 continuous glucose monitoring.
10.1177_0963689720947098-fig1.jpg
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There was no time spent in hypoglycemia comparing to 2% before islet graft (Fig. 1B).
The "old school" purification schema.
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Transplant Wash Medium: CMRL 1066 (Mediatech) with 2% of human serum albumin (Baxter), 1 M of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES; Mediatech), and 10 U/ml of heparin (Polfa Warszawa S.A.) and again allowed to settle for 5 min. Clear supernatant from the above of the sedimenting tissue was collected and discarded, and the tissue pellet was immediately segregated using a 10 ml pipet as follows: the first 7 ml was collected from the surface of the pellet and transferred to separate 250 ml conical tubes (Corning), the next 7 ml from the middle portion and transferred to separate 250 ml conical tubes (Corning), and the bottom portion was left in the same conical tube. The schema of islet purification by sedimentation at gravity unit is shown in Fig. 2.
The pellet volumes and IEQ for each fraction during the initial and supplemental purification. IEQ: islet equivalent.
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The pellet volumes and IEQ for each fraction during the supplemental purification are presented in Fig. 3. Islets from the post-isolation high-purity fraction (0.6 ml) were collected from T-175 flasks (Thermo Fisher Scientific) into a 250 ml conical tube (Corning) and washed three times with CIT Transplant Wash Medium (Fig. 3). Individual fractions with the highest IEQ per milliliter pellet volume were selected from the post-isolation fractions with purity >30% for the final product transplantation (2 ml from the upper and 4 ml from the middle-purity fraction) (Fig. 3). Transplant Medium: CMRL 1066 (Mediatech) supplemented with 2% human serum albumin (Baxter) and 1 M HEPES (Mediatech) and prepared in three infusion bags (Transfer Pack Container, Fenwal, Lake Zurich, IL, USA) as follows: post-isolation high-purity islets (0.6 ml) and the upper fraction of tissue retrieved from the post-isolation 20% islet purity were combined and prepared in the first bag (beige rectangles in Fig. 3). The middle fraction of the tissue from the post-isolation 20% islet purity fraction was prepared in the second bag (blue rectangle in Fig. 3) and the bottom portion of the tissue from the post-isolation 10% islet purity fraction was prepared in the third bag (green rectangle in Fig. 3).