Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical Cord
Recent studies have demonstrated that the umbilical cord (UC) is an excellent source of mesenchymal stromal cells (MSCs). However, current protocols for extracting and culturing UC-MSCs do not meet current good manufacturing practice (cGMP) standards, in part due to the use of xenogeneic reagents. T...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2016-01-01
|
| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2016/3274054 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832548331074617344 |
|---|---|
| author | Ryan J. Emnett Aparna Kaul Aleksandar Babic Vicki Geiler Donna Regan Gilad Gross Salem Akel |
| author_facet | Ryan J. Emnett Aparna Kaul Aleksandar Babic Vicki Geiler Donna Regan Gilad Gross Salem Akel |
| author_sort | Ryan J. Emnett |
| collection | DOAJ |
| description | Recent studies have demonstrated that the umbilical cord (UC) is an excellent source of mesenchymal stromal cells (MSCs). However, current protocols for extracting and culturing UC-MSCs do not meet current good manufacturing practice (cGMP) standards, in part due to the use of xenogeneic reagents. To support the development of a cGMP-compliant method, we have examined an enzyme-free isolation method utilizing tissue homogenization (t-H) followed by culture in human platelet lysate (PL) supplemented media. The yield and viability of cells after t-H were comparable to those obtained after collagenase digestion (Col-D). Importantly, kinetic analysis of cultured cells showed logarithmic growth over 10 tested passages, although the rate of cell division was lower for t-H as compared to Col-D. This slower growth of t-H-derived cells was also reflected in their longer population doubling time. Interestingly, there was no difference in the expression of mesenchymal markers and trilineage differentiation potential of cells generated using either method. Finally, t-H-derived cells had greater clonogenic potential compared to Col-D/FBS but not Col-D/PL and were able to maintain CFU-F capacity through P7. This bench scale study demonstrates the possibility of generating therapeutic doses of good quality UC-MSCs within a reasonable length of time using t-H and PL. |
| format | Article |
| id | doaj-art-6581fcae41ea4a71a7c76aa055b2184c |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-6581fcae41ea4a71a7c76aa055b2184c2025-02-03T06:14:18ZengWileyStem Cells International1687-966X1687-96782016-01-01201610.1155/2016/32740543274054Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical CordRyan J. Emnett0Aparna Kaul1Aleksandar Babic2Vicki Geiler3Donna Regan4Gilad Gross5Salem Akel6St. Louis Cord Blood Bank/Cellular Therapy Laboratory, SSM Health Cardinal Glennon Children’s Hospital, St. Louis, MO 63110, USASt. Louis Cord Blood Bank/Cellular Therapy Laboratory, SSM Health Cardinal Glennon Children’s Hospital, St. Louis, MO 63110, USASt. Louis Cord Blood Bank/Cellular Therapy Laboratory, SSM Health Cardinal Glennon Children’s Hospital, St. Louis, MO 63110, USASt. Louis Cord Blood Bank/Cellular Therapy Laboratory, SSM Health Cardinal Glennon Children’s Hospital, St. Louis, MO 63110, USASt. Louis Cord Blood Bank/Cellular Therapy Laboratory, SSM Health Cardinal Glennon Children’s Hospital, St. Louis, MO 63110, USASSM Health St. Mary’s Hospital, St. Louis, MO 63117, USASt. Louis Cord Blood Bank/Cellular Therapy Laboratory, SSM Health Cardinal Glennon Children’s Hospital, St. Louis, MO 63110, USARecent studies have demonstrated that the umbilical cord (UC) is an excellent source of mesenchymal stromal cells (MSCs). However, current protocols for extracting and culturing UC-MSCs do not meet current good manufacturing practice (cGMP) standards, in part due to the use of xenogeneic reagents. To support the development of a cGMP-compliant method, we have examined an enzyme-free isolation method utilizing tissue homogenization (t-H) followed by culture in human platelet lysate (PL) supplemented media. The yield and viability of cells after t-H were comparable to those obtained after collagenase digestion (Col-D). Importantly, kinetic analysis of cultured cells showed logarithmic growth over 10 tested passages, although the rate of cell division was lower for t-H as compared to Col-D. This slower growth of t-H-derived cells was also reflected in their longer population doubling time. Interestingly, there was no difference in the expression of mesenchymal markers and trilineage differentiation potential of cells generated using either method. Finally, t-H-derived cells had greater clonogenic potential compared to Col-D/FBS but not Col-D/PL and were able to maintain CFU-F capacity through P7. This bench scale study demonstrates the possibility of generating therapeutic doses of good quality UC-MSCs within a reasonable length of time using t-H and PL.http://dx.doi.org/10.1155/2016/3274054 |
| spellingShingle | Ryan J. Emnett Aparna Kaul Aleksandar Babic Vicki Geiler Donna Regan Gilad Gross Salem Akel Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical Cord Stem Cells International |
| title | Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical Cord |
| title_full | Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical Cord |
| title_fullStr | Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical Cord |
| title_full_unstemmed | Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical Cord |
| title_short | Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical Cord |
| title_sort | evaluation of tissue homogenization to support the generation of gmp compliant mesenchymal stromal cells from the umbilical cord |
| url | http://dx.doi.org/10.1155/2016/3274054 |
| work_keys_str_mv | AT ryanjemnett evaluationoftissuehomogenizationtosupportthegenerationofgmpcompliantmesenchymalstromalcellsfromtheumbilicalcord AT aparnakaul evaluationoftissuehomogenizationtosupportthegenerationofgmpcompliantmesenchymalstromalcellsfromtheumbilicalcord AT aleksandarbabic evaluationoftissuehomogenizationtosupportthegenerationofgmpcompliantmesenchymalstromalcellsfromtheumbilicalcord AT vickigeiler evaluationoftissuehomogenizationtosupportthegenerationofgmpcompliantmesenchymalstromalcellsfromtheumbilicalcord AT donnaregan evaluationoftissuehomogenizationtosupportthegenerationofgmpcompliantmesenchymalstromalcellsfromtheumbilicalcord AT giladgross evaluationoftissuehomogenizationtosupportthegenerationofgmpcompliantmesenchymalstromalcellsfromtheumbilicalcord AT salemakel evaluationoftissuehomogenizationtosupportthegenerationofgmpcompliantmesenchymalstromalcellsfromtheumbilicalcord |