{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/110709"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/110709","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Single-cell analysis of hematopoietic cell fate on niche-mimetic platforms using Raman microspectroscopy","abstract":"Regenerative therapies are predicated on our ability to achieve controlled and reproducible expansion of stem cells ex vivo. Hematopoietic stem cells (HSCs) are the best-studied class of tissue-restricted stem cells and their microenvironment in vivo has also been the subject of considerable research. Although a host of cellular, molecular and physical cues that regulate stem cell fate in vivo have been identified, efforts to culture HSCs for extended periods by recapitulating some of these interactions in vitro have met with limited success. Miniaturized biomaterial platforms that present tens to hundreds of putative niche components can help screen individual factors or their combinations which drive quiescence, self-renewal and differentiation in vitro while reducing the numbers of rare cell populations required for testing. Screening approaches rely on accurate identification of specific stem cell outcomes in order to correlate extrinsic cues with the fates they elicit. Traditionally, HSCs and their progeny populations have been identified using time-consuming functional assays or immunofluorescence detection of a combination of surface antigens, both of which are invasive and preclude longitudinal analysis of cell fate in culture. Methods that allow rapid, label-free and location-specific characterization of the differentiation stages of individual cells would enable the rapid development of concurrent screens of the effects of niche components and their combinations on stem cell fate specification. Raman spectroscopy has emerged as one of the few analytical approaches that can be used to noninvasively assay the biomolecular composition of cells. Combining Raman spectroscopy with multivariate methods such as partial least-squares discriminant analysis and self-organizing maps may enable objective classification of specific stem cell outcomes on screening platforms, or facilitate investigations into the clonal heterogeneity of stem cells, respectively. The work presented herein describes the use of spontaneous Raman microspectroscopy and multivariate methods as a noninvasive technique for following hematopoietic stem cells and their progeny in vitro. Chapter 1 introduces the field of tissue engineering and the role of the stem cell microenvironment in guiding stem cell fate especially as it relates to the hematopoietic stem cell and lays down the background and motivation for label-free analytical approaches for phenotypic analysis of stem cells in vitro. Chapter 2 discusses the development of a hydrogel culture platform that is compatible with Raman spectroscopy, and demonstrates the capability of multivariate methods to capture subtle changes in the biochemical make-up of cells using mammalian cell lines. Chapter 3 uses a monocytic leukemia cell line capable of differentiating into macrophages (THP-1 line) to build on the work presented in Chapter 2 and provide proof of concept for the feasibility of real-time tracking of cell differentiation using Raman spectroscopy. This chapter introduces self-organizing maps (SOMs). Finally, Chapter 4 shows the successful use of Raman spectroscopy and partial least-squares discriminant analysis for determining the differentiation stages of individual living hematopoietic stem cells and their progeny on biomaterial substrates with varying stiffness. Chapter 5 presents a short discussion of the implications of this work for the field and summarizes some of the issues and challenges that must be addressed in future work using Raman spectroscopy on HSCs.","abstract_html":"Regenerative therapies are predicated on our ability to achieve controlled and reproducible expansion of stem cells ex vivo. Hematopoietic stem cells (HSCs) are the best-studied class of tissue-restricted stem cells and their microenvironment in vivo has also been the subject of considerable research. Although a host of cellular, molecular and physical cues that regulate stem cell fate in vivo have been identified, efforts to culture HSCs for extended periods by recapitulating some of these interactions in vitro have met with limited success. Miniaturized biomaterial platforms that present tens to hundreds of putative niche components can help screen individual factors or their combinations which drive quiescence, self-renewal and differentiation in vitro while reducing the numbers of rare cell populations required for testing. Screening approaches rely on accurate identification of specific stem cell outcomes in order to correlate extrinsic cues with the fates they elicit. Traditionally, HSCs and their progeny populations have been identified using time-consuming functional assays or immunofluorescence detection of a combination of surface antigens, both of which are invasive and preclude longitudinal analysis of cell fate in culture. Methods that allow rapid, label-free and location-specific characterization of the differentiation stages of individual cells would enable the rapid development of concurrent screens of the effects of niche components and their combinations on stem cell fate specification. Raman spectroscopy has emerged as one of the few analytical approaches that can be used to noninvasively assay the biomolecular composition of cells. Combining Raman spectroscopy with multivariate methods such as partial least-squares discriminant analysis and self-organizing maps may enable objective classification of specific stem cell outcomes on screening platforms, or facilitate investigations into the clonal heterogeneity of stem cells, respectively. The work presented herein describes the use of spontaneous Raman microspectroscopy and multivariate methods as a noninvasive technique for following hematopoietic stem cells and their progeny in vitro. Chapter 1 introduces the field of tissue engineering and the role of the stem cell microenvironment in guiding stem cell fate especially as it relates to the hematopoietic stem cell and lays down the background and motivation for label-free analytical approaches for phenotypic analysis of stem cells in vitro. Chapter 2 discusses the development of a hydrogel culture platform that is compatible with Raman spectroscopy, and demonstrates the capability of multivariate methods to capture subtle changes in the biochemical make-up of cells using mammalian cell lines. Chapter 3 uses a monocytic leukemia cell line capable of differentiating into macrophages (THP-1 line) to build on the work presented in Chapter 2 and provide proof of concept for the feasibility of real-time tracking of cell differentiation using Raman spectroscopy. This chapter introduces self-organizing maps (SOMs). Finally, Chapter 4 shows the successful use of Raman spectroscopy and partial least-squares discriminant analysis for determining the differentiation stages of individual living hematopoietic stem cells and their progeny on biomaterial substrates with varying stiffness. Chapter 5 presents a short discussion of the implications of this work for the field and summarizes some of the issues and challenges that must be addressed in future work using Raman spectroscopy on HSCs.","abstract_has_math":false,"creators":["Majumdar, Sayani"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Chemical Engineering","degree_department":null,"school":null,"contributors":["Kraft, Mary L","Harley, Brendan A","Kong, Hyun Joon","Underhill, Gregory H"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2021,"date_issued":"2021-09-17T02:34:40Z","date_published":"2021-09-17T02:34:40Z","updated_at":"2026-07-22T22:24:52Z","subjects":["Raman microspectroscopy","Multivariate analysis","Cell discrimination","Single-cell Raman spectra","Hematopoietic stem cells","Stem cell differentiation","Noninvasive phenotypic analysis","Stem cell niche"],"languages":["en"],"rights":["Copyright 2021 Sayani Majumdar"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/110709","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Kraft, Mary L","Harley, Brendan A","Kong, Hyun Joon","Underhill, Gregory H"]},{"key":"dc:creator","label":"Author","values":["Majumdar, Sayani"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2021-09-17T02:34:40Z","2023-09-17T02:34:57Z","2021-04-22","2021-05"]},{"key":"dc:type","label":"Dc Type","values":["text","Thesis"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemical Engineering"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Raman microspectroscopy","Multivariate analysis","Cell discrimination","Single-cell Raman spectra","Hematopoietic stem cells","Stem cell differentiation","Noninvasive phenotypic analysis","Stem cell niche"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2021 Sayani Majumdar"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/110709"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Regenerative therapies are predicated on our ability to achieve controlled and reproducible expansion of stem cells ex vivo. Hematopoietic stem cells (HSCs) are the best-studied class of tissue-restricted stem cells and their microenvironment in vivo has also been the subject of considerable research. Although a host of cellular, molecular and physical cues that regulate stem cell fate in vivo have been identified, efforts to culture HSCs for extended periods by recapitulating some of these interactions in vitro have met with limited success. Miniaturized biomaterial platforms that present tens to hundreds of putative niche components can help screen individual factors or their combinations which drive quiescence, self-renewal and differentiation in vitro while reducing the numbers of rare cell populations required for testing. Screening approaches rely on accurate identification of specific stem cell outcomes in order to correlate extrinsic cues with the fates they elicit. Traditionally, HSCs and their progeny populations have been identified using time-consuming functional assays or immunofluorescence detection of a combination of surface antigens, both of which are invasive and preclude longitudinal analysis of cell fate in culture. Methods that allow rapid, label-free and location-specific characterization of the differentiation stages of individual cells would enable the rapid development of concurrent screens of the effects of niche components and their combinations on stem cell fate specification. Raman spectroscopy has emerged as one of the few analytical approaches that can be used to noninvasively assay the biomolecular composition of cells. Combining Raman spectroscopy with multivariate methods such as partial least-squares discriminant analysis and self-organizing maps may enable objective classification of specific stem cell outcomes on screening platforms, or facilitate investigations into the clonal heterogeneity of stem cells, respectively. The work presented herein describes the use of spontaneous Raman microspectroscopy and multivariate methods as a noninvasive technique for following hematopoietic stem cells and their progeny in vitro. Chapter 1 introduces the field of tissue engineering and the role of the stem cell microenvironment in guiding stem cell fate especially as it relates to the hematopoietic stem cell and lays down the background and motivation for label-free analytical approaches for phenotypic analysis of stem cells in vitro. Chapter 2 discusses the development of a hydrogel culture platform that is compatible with Raman spectroscopy, and demonstrates the capability of multivariate methods to capture subtle changes in the biochemical make-up of cells using mammalian cell lines. Chapter 3 uses a monocytic leukemia cell line capable of differentiating into macrophages (THP-1 line) to build on the work presented in Chapter 2 and provide proof of concept for the feasibility of real-time tracking of cell differentiation using Raman spectroscopy. This chapter introduces self-organizing maps (SOMs). Finally, Chapter 4 shows the successful use of Raman spectroscopy and partial least-squares discriminant analysis for determining the differentiation stages of individual living hematopoietic stem cells and their progeny on biomaterial substrates with varying stiffness. Chapter 5 presents a short discussion of the implications of this work for the field and summarizes some of the issues and challenges that must be addressed in future work using Raman spectroscopy on HSCs.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2023-05-01","The student, Sayani Majumdar, accepted the attached license on 2021-04-21 at 02:52.","The student, Sayani Majumdar, submitted this Dissertation for approval on 2021-04-21 at 03:13.","This Dissertation was approved for publication on 2021-04-22 at 13:43.","DSpace SAF Submission Ingestion Package generated from Vireo submission #16456 on 2021-09-16 at 17:04:12","Made available in DSpace on 2021-09-17T02:34:40Z (GMT). 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Hematopoietic stem cells (HSCs) are the best-studied class of tissue-restricted stem cells and their microenvironment in vivo has also been the subject of considerable research. Although a host of cellular, molecular and physical cues that regulate stem cell fate in vivo have been identified, efforts to culture HSCs for extended periods by recapitulating some of these interactions in vitro have met with limited success. Miniaturized biomaterial platforms that present tens to hundreds of putative niche components can help screen individual factors or their combinations which drive quiescence, self-renewal and differentiation in vitro while reducing the numbers of rare cell populations required for testing. Screening approaches rely on accurate identification of specific stem cell outcomes in order to correlate extrinsic cues with the fates they elicit. Traditionally, HSCs and their progeny populations have been identified using time-consuming functional assays or immunofluorescence detection of a combination of surface antigens, both of which are invasive and preclude longitudinal analysis of cell fate in culture. Methods that allow rapid, label-free and location-specific characterization of the differentiation stages of individual cells would enable the rapid development of concurrent screens of the effects of niche components and their combinations on stem cell fate specification. Raman spectroscopy has emerged as one of the few analytical approaches that can be used to noninvasively assay the biomolecular composition of cells. Combining Raman spectroscopy with multivariate methods such as partial least-squares discriminant analysis and self-organizing maps may enable objective classification of specific stem cell outcomes on screening platforms, or facilitate investigations into the clonal heterogeneity of stem cells, respectively. The work presented herein describes the use of spontaneous Raman microspectroscopy and multivariate methods as a noninvasive technique for following hematopoietic stem cells and their progeny in vitro. Chapter 1 introduces the field of tissue engineering and the role of the stem cell microenvironment in guiding stem cell fate especially as it relates to the hematopoietic stem cell and lays down the background and motivation for label-free analytical approaches for phenotypic analysis of stem cells in vitro. Chapter 2 discusses the development of a hydrogel culture platform that is compatible with Raman spectroscopy, and demonstrates the capability of multivariate methods to capture subtle changes in the biochemical make-up of cells using mammalian cell lines. Chapter 3 uses a monocytic leukemia cell line capable of differentiating into macrophages (THP-1 line) to build on the work presented in Chapter 2 and provide proof of concept for the feasibility of real-time tracking of cell differentiation using Raman spectroscopy. This chapter introduces self-organizing maps (SOMs). Finally, Chapter 4 shows the successful use of Raman spectroscopy and partial least-squares discriminant analysis for determining the differentiation stages of individual living hematopoietic stem cells and their progeny on biomaterial substrates with varying stiffness. Chapter 5 presents a short discussion of the implications of this work for the field and summarizes some of the issues and challenges that must be addressed in future work using Raman spectroscopy on HSCs.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2023-05-01","The student, Sayani Majumdar, accepted the attached license on 2021-04-21 at 02:52.","The student, Sayani Majumdar, submitted this Dissertation for approval on 2021-04-21 at 03:13.","This Dissertation was approved for publication on 2021-04-22 at 13:43.","DSpace SAF Submission Ingestion Package generated from Vireo submission #16456 on 2021-09-16 at 17:04:12","Made available in DSpace on 2021-09-17T02:34:40Z (GMT). No. of bitstreams: 3 MAJUMDAR-DISSERTATION-2021.pdf: 4556444 bytes, checksum: 6cdd04783e76d6bac86431086669ffcd (MD5) Copyright E Form_BIP20.pdf: 397364 bytes, checksum: 7bbd1259c5c71628a50207392765fceb (MD5) LICENSE.txt: 4212 bytes, checksum: c1c3e9bff330a33a94fda188210d0490 (MD5) Previous issue date: 2021-04-22","Embargo set by: Seth Robbins for item 118552 Lift date: 2023-09-17T02:34:57Z Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","U of I Only"],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/2142/110709"],"dc:language":["en"],"dc:rights":["Copyright 2021 Sayani Majumdar"],"dc:subject":["Raman microspectroscopy","Multivariate analysis","Cell discrimination","Single-cell Raman spectra","Hematopoietic stem cells","Stem cell differentiation","Noninvasive phenotypic analysis","Stem cell niche"],"dc:title":["Single-cell analysis of hematopoietic cell fate on niche-mimetic platforms using Raman microspectroscopy"],"dc:type":["text","Thesis"],"thesis:degree_discipline":["Chemical Engineering"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:24:52Z"}