Expansion of human umbilical cord derived mesenchymal stem cells in regenerative medicine
Stem cells are undifferentiated cells that possess the potential for self-renewal with the capacity to differentiate into multiple lineages. In humans, their limited numbers pose a challenge in fulfilling the necessary demands for the regeneration and repair of damaged tissues or organs. Studies sug...
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Veröffentlicht in: | World journal of stem cells 2024-04, Vol.16 (4), p.410-433 |
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description | Stem cells are undifferentiated cells that possess the potential for self-renewal with the capacity to differentiate into multiple lineages. In humans, their limited numbers pose a challenge in fulfilling the necessary demands for the regeneration and repair of damaged tissues or organs. Studies suggested that mesenchymal stem cells (MSCs), necessary for repair and regeneration via transplantation, require doses ranging from 10 to 400 million cells. Furthermore, the limited expansion of MSCs restricts their therapeutic application.
To optimize a novel protocol to achieve qualitative and quantitative expansion of MSCs to reach the targeted number of cells for cellular transplantation and minimize the limitations in stem cell therapy protocols.
Human umbilical cord (hUC) tissue derived MSCs were obtained and re-cultured. These cultured cells were subjected to the following evaluation procedures: Immunophenotyping, immunocytochemical staining, trilineage differentiation, population doubling time and number, gene expression markers for proliferation, cell cycle progression, senescence-associated β-galactosidase assay, human telomerase reverse transcriptase (hTERT) expression, mycoplasma, cytomegalovirus and endotoxin detection.
Analysis of pluripotent gene markers
,
, and
in recultured hUC-MSC revealed no significant differences. The immunophenotypic markers CD90, CD73, CD105, CD44, vimentin, CD29, Stro-1, and Lin28 were positively expressed by these recultured expanded MSCs, and were found negative for CD34, CD11b, CD19, CD45, and HLA-DR. The recultured hUC-MSC population continued to expand through passage 15. Proliferative gene expression of
,
, and
showed no significant variation between recultured hUC-MSC groups. Nevertheless, a significant increase (
< 0.001) in the mitotic phase of the cell cycle was observed in recultured hUC-MSCs. Cellular senescence markers (hTERT expression and β-galactosidase activity) did not show any negative effect on recultured hUC-MSCs. Additionally, quality control assessments consistently confirmed the absence of mycoplasma, cytomegalovirus, and endotoxin contamination.
This study proposes the development of a novel protocol for efficiently expanding stem cell population. This would address the growing demand for larger stem cell doses needed for cellular transplantation and will significantly improve the feasibility of stem cell based therapies. |
doi_str_mv | 10.4252/wjsc.v16.i4.410 |
format | Article |
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To optimize a novel protocol to achieve qualitative and quantitative expansion of MSCs to reach the targeted number of cells for cellular transplantation and minimize the limitations in stem cell therapy protocols.
Human umbilical cord (hUC) tissue derived MSCs were obtained and re-cultured. These cultured cells were subjected to the following evaluation procedures: Immunophenotyping, immunocytochemical staining, trilineage differentiation, population doubling time and number, gene expression markers for proliferation, cell cycle progression, senescence-associated β-galactosidase assay, human telomerase reverse transcriptase (hTERT) expression, mycoplasma, cytomegalovirus and endotoxin detection.
Analysis of pluripotent gene markers
,
, and
in recultured hUC-MSC revealed no significant differences. The immunophenotypic markers CD90, CD73, CD105, CD44, vimentin, CD29, Stro-1, and Lin28 were positively expressed by these recultured expanded MSCs, and were found negative for CD34, CD11b, CD19, CD45, and HLA-DR. The recultured hUC-MSC population continued to expand through passage 15. Proliferative gene expression of
,
, and
showed no significant variation between recultured hUC-MSC groups. Nevertheless, a significant increase (
< 0.001) in the mitotic phase of the cell cycle was observed in recultured hUC-MSCs. Cellular senescence markers (hTERT expression and β-galactosidase activity) did not show any negative effect on recultured hUC-MSCs. Additionally, quality control assessments consistently confirmed the absence of mycoplasma, cytomegalovirus, and endotoxin contamination.
This study proposes the development of a novel protocol for efficiently expanding stem cell population. This would address the growing demand for larger stem cell doses needed for cellular transplantation and will significantly improve the feasibility of stem cell based therapies.</description><identifier>ISSN: 1948-0210</identifier><identifier>EISSN: 1948-0210</identifier><identifier>DOI: 10.4252/wjsc.v16.i4.410</identifier><identifier>PMID: 38690517</identifier><language>eng</language><publisher>United States: Baishideng Publishing Group Inc</publisher><subject>Basic Study</subject><ispartof>World journal of stem cells, 2024-04, Vol.16 (4), p.410-433</ispartof><rights>The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.</rights><rights>The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved. 2024</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c283t-5435d6e1412afac26de01a8591688d441ac78dad466a8d6b97c20422d6a11aea3</citedby><cites>FETCH-LOGICAL-c283t-5435d6e1412afac26de01a8591688d441ac78dad466a8d6b97c20422d6a11aea3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC11056638/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC11056638/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38690517$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rajput, Shafiqa Naeem</creatorcontrib><creatorcontrib>Naeem, Bushra Kiran</creatorcontrib><creatorcontrib>Ali, Anwar</creatorcontrib><creatorcontrib>Salim, Asmat</creatorcontrib><creatorcontrib>Khan, Irfan</creatorcontrib><title>Expansion of human umbilical cord derived mesenchymal stem cells in regenerative medicine</title><title>World journal of stem cells</title><addtitle>World J Stem Cells</addtitle><description>Stem cells are undifferentiated cells that possess the potential for self-renewal with the capacity to differentiate into multiple lineages. In humans, their limited numbers pose a challenge in fulfilling the necessary demands for the regeneration and repair of damaged tissues or organs. Studies suggested that mesenchymal stem cells (MSCs), necessary for repair and regeneration via transplantation, require doses ranging from 10 to 400 million cells. Furthermore, the limited expansion of MSCs restricts their therapeutic application.
To optimize a novel protocol to achieve qualitative and quantitative expansion of MSCs to reach the targeted number of cells for cellular transplantation and minimize the limitations in stem cell therapy protocols.
Human umbilical cord (hUC) tissue derived MSCs were obtained and re-cultured. These cultured cells were subjected to the following evaluation procedures: Immunophenotyping, immunocytochemical staining, trilineage differentiation, population doubling time and number, gene expression markers for proliferation, cell cycle progression, senescence-associated β-galactosidase assay, human telomerase reverse transcriptase (hTERT) expression, mycoplasma, cytomegalovirus and endotoxin detection.
Analysis of pluripotent gene markers
,
, and
in recultured hUC-MSC revealed no significant differences. The immunophenotypic markers CD90, CD73, CD105, CD44, vimentin, CD29, Stro-1, and Lin28 were positively expressed by these recultured expanded MSCs, and were found negative for CD34, CD11b, CD19, CD45, and HLA-DR. The recultured hUC-MSC population continued to expand through passage 15. Proliferative gene expression of
,
, and
showed no significant variation between recultured hUC-MSC groups. Nevertheless, a significant increase (
< 0.001) in the mitotic phase of the cell cycle was observed in recultured hUC-MSCs. Cellular senescence markers (hTERT expression and β-galactosidase activity) did not show any negative effect on recultured hUC-MSCs. Additionally, quality control assessments consistently confirmed the absence of mycoplasma, cytomegalovirus, and endotoxin contamination.
This study proposes the development of a novel protocol for efficiently expanding stem cell population. This would address the growing demand for larger stem cell doses needed for cellular transplantation and will significantly improve the feasibility of stem cell based therapies.</description><subject>Basic Study</subject><issn>1948-0210</issn><issn>1948-0210</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><recordid>eNpVkUFvGyEQRlHVqIlcn3urOOZih2FZzJ6iyEqTSpZ6aQ85oTGMY6Jd1oFdt_n3wXISuVxAmsc3A4-xbyDmStby6u9TdvM96HlQcwXiE7uARpmZkCA-n5zP2TTnJ1GWqrVW8gs7r4xuRA2LC_Zw-2-HMYc-8n7Dt2OHkY_dOrTBYctdnzz3lMKePO8oU3Tbl64U8kAdd9S2mYfIEz1SpIRD4QrmgwuRvrKzDbaZpm_7hP35cft7eT9b_br7ubxZzZw01TCrVVV7TaBA4gad1J4EoKkb0MZ4pQDdwnj0Sms0Xq-bhZNCSek1AiBhNWHXx9zduC69HcUhYWt3KXSYXmyPwf5fiWFrH_u9BRDlPypTEi7fElL_PFIebBfy4XEYqR-zrYRqFqDrShX06oi61OecaPPRB4Q9SLEHKbZIsUHZIqXc-H463gf_rqB6Bd07i3Y</recordid><startdate>20240426</startdate><enddate>20240426</enddate><creator>Rajput, Shafiqa Naeem</creator><creator>Naeem, Bushra Kiran</creator><creator>Ali, Anwar</creator><creator>Salim, Asmat</creator><creator>Khan, Irfan</creator><general>Baishideng Publishing Group Inc</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20240426</creationdate><title>Expansion of human umbilical cord derived mesenchymal stem cells in regenerative medicine</title><author>Rajput, Shafiqa Naeem ; Naeem, Bushra Kiran ; Ali, Anwar ; Salim, Asmat ; Khan, Irfan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c283t-5435d6e1412afac26de01a8591688d441ac78dad466a8d6b97c20422d6a11aea3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Basic Study</topic><toplevel>online_resources</toplevel><creatorcontrib>Rajput, Shafiqa Naeem</creatorcontrib><creatorcontrib>Naeem, Bushra Kiran</creatorcontrib><creatorcontrib>Ali, Anwar</creatorcontrib><creatorcontrib>Salim, Asmat</creatorcontrib><creatorcontrib>Khan, Irfan</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>World journal of stem cells</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rajput, Shafiqa Naeem</au><au>Naeem, Bushra Kiran</au><au>Ali, Anwar</au><au>Salim, Asmat</au><au>Khan, Irfan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expansion of human umbilical cord derived mesenchymal stem cells in regenerative medicine</atitle><jtitle>World journal of stem cells</jtitle><addtitle>World J Stem Cells</addtitle><date>2024-04-26</date><risdate>2024</risdate><volume>16</volume><issue>4</issue><spage>410</spage><epage>433</epage><pages>410-433</pages><issn>1948-0210</issn><eissn>1948-0210</eissn><abstract>Stem cells are undifferentiated cells that possess the potential for self-renewal with the capacity to differentiate into multiple lineages. In humans, their limited numbers pose a challenge in fulfilling the necessary demands for the regeneration and repair of damaged tissues or organs. Studies suggested that mesenchymal stem cells (MSCs), necessary for repair and regeneration via transplantation, require doses ranging from 10 to 400 million cells. Furthermore, the limited expansion of MSCs restricts their therapeutic application.
To optimize a novel protocol to achieve qualitative and quantitative expansion of MSCs to reach the targeted number of cells for cellular transplantation and minimize the limitations in stem cell therapy protocols.
Human umbilical cord (hUC) tissue derived MSCs were obtained and re-cultured. These cultured cells were subjected to the following evaluation procedures: Immunophenotyping, immunocytochemical staining, trilineage differentiation, population doubling time and number, gene expression markers for proliferation, cell cycle progression, senescence-associated β-galactosidase assay, human telomerase reverse transcriptase (hTERT) expression, mycoplasma, cytomegalovirus and endotoxin detection.
Analysis of pluripotent gene markers
,
, and
in recultured hUC-MSC revealed no significant differences. The immunophenotypic markers CD90, CD73, CD105, CD44, vimentin, CD29, Stro-1, and Lin28 were positively expressed by these recultured expanded MSCs, and were found negative for CD34, CD11b, CD19, CD45, and HLA-DR. The recultured hUC-MSC population continued to expand through passage 15. Proliferative gene expression of
,
, and
showed no significant variation between recultured hUC-MSC groups. Nevertheless, a significant increase (
< 0.001) in the mitotic phase of the cell cycle was observed in recultured hUC-MSCs. Cellular senescence markers (hTERT expression and β-galactosidase activity) did not show any negative effect on recultured hUC-MSCs. Additionally, quality control assessments consistently confirmed the absence of mycoplasma, cytomegalovirus, and endotoxin contamination.
This study proposes the development of a novel protocol for efficiently expanding stem cell population. This would address the growing demand for larger stem cell doses needed for cellular transplantation and will significantly improve the feasibility of stem cell based therapies.</abstract><cop>United States</cop><pub>Baishideng Publishing Group Inc</pub><pmid>38690517</pmid><doi>10.4252/wjsc.v16.i4.410</doi><tpages>24</tpages><oa>free_for_read</oa></addata></record> |
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title | Expansion of human umbilical cord derived mesenchymal stem cells in regenerative medicine |
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