Macroencapsulated secretory cells
一种胶原分泌、细胞的技术，应用在细胞胶囊化、微型胶囊、胶囊输送等方向，能够解决膜功能疑问等问题
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Embodiment 1
[0045] Example 1 Isolation of pancreatic islets
[0046] Islets were isolated from rats using a modification of the method described by Gotoh, et. al. Transplantation 40:437 (1985).
[0047] Collagen solution (XI type collagenase, Sigma Chemical, St.Louis, MO, 1mg / ml, containing 2mg / ml Sigma V type bovine serum albumin and 1mg / ml CaCl 2 ) into the pancreas via the common bile canaliculus (Gotoh et al, Transplantation 40:437 (1985)). Remove the pancreas and collect in a flask on ice. Once the pancreases from the 4 rats have been collected, place the flasks in a 38°C water bath for 30 minutes. The resulting digested tissue was washed 4 times with cold (8°C) HBSS (Hank's Balanced Salt Solution).
[0048] Undigested tissue, large lymph nodes, and other extraneous matter are removed by repeated flow through the tissue with removal of the supernatant. Purified islets were separated under a discontinuous Ficoll gradient consisting of 25%, 23%, 21% and 11% Ficoll layers prepared i...
Embodiment 2
[0052] A Preparation of agarose-coated agarose-collagen islet macrobeads
[0053] The 1000 islets obtained by the method of Example 1 were washed 4 times in RPMI complete medium with fetal bovine serum removed. The islets were then added to a test tube containing 50 µl of 1% atelocollagen solution in phosphate buffered saline to suspend the islets. 100 μl of 1% low-viscosity agarose (SigmaType XII) prepared in RPMI or MEN (Minimal Essential Medium) maintained at 60°C was added to the collagen-islet suspension, and the contents of the tube were immediately separated as individual Large drops were transferred onto sterile mineral oil or Teflon sheets kept at room temperature. After 1 minute, the drops turned into semi-solid beads, which were then transferred to RPMI antibiotic medium at 37°C, and the beads were washed 3 times with the same medium to remove all oil. Finally, rinse twice with complete medium (37°C) in a humidified medium containing 5% CO at 37°C 2 Incubate over...
Embodiment 3
[0061] Example 3 Transplantation of islet beads into mice
[0062] A Recipient and donor rats
[0063] The mice used were male C57BL / 6 and BALB / c species, and recipient mice were rendered diabetic with a single intravenous injection of streptozotocin (170-200 mg / kg).
[0064] Non-fasting plasma glucose levels were measured prior to induction of diabetes and blood glucose levels were monitored in all recipient mice by tail vein blood samples using an ExacTech Pen Senson, only those mice with serum glucose levels > 400 mg / dl on the day of transplantation were used.
[0065] Wistar Furth rats were used as xenograft donors.
[0066] B Xenotransplantation of large islet beads into the peritoneal cavity
[0067] At the time of xenotransplantation, gently place the islet beads obtained in Example 2 (A), (B), and (C) into respective plates containing RPMI antibiotic medium, and replace the culture medium to remove all serum proteins. Base 3 times, diabetic recipient mice were anest...
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