Endothelial-neurosphere crosstalk in microwell arrays regulates self-renewal and differentiation of human neural stem cells

Kisuk Yang; Jong Seung Lee; Sewoon Han; Yoonhee Jin; Ann-Na Cho; Gyeong-Eon Chang; Eunji Cheong; Ji Hun Yang; Seok Chung; Seung-Woo Cho

Journal of Industrial and Engineering Chemistry, Vol.74, 148-157, June, 2019

DOI10.1016/j.jiec.2019.02.021 Export Citation

Endothelial-neurosphere crosstalk in microwell arrays regulates self-renewal and differentiation of human neural stem cells

Kisuk Yang¹, Jong Seung Lee¹, Sewoon Han², Yoonhee Jin¹, Ann-Na Cho¹, Gyeong-Eon Chang¹, Eunji Cheong¹, Ji Hun Yang², Seok Chung^{2, 3, †}, and Seung-Woo Cho^{1, †}

¹Department of Biotechnology, Yonsei University, Seoul 03722, Republic of Korea
²Department of Mechanical Engineering, Korea University, Seoul 02841, Republic of Korea
³KU-KIST Graduate School of Converging Science and Technology, Republic of Korea

E-mail:,

Control of neural stem cell (NSC) self-renewal and differentiationis of great importance to improve its therapeutic efficacy in the treatment of neurodegenerative diseases. Neurosphere culture for NSC expansion under undifferentiation condition determines the self-renewal capacity and differentiation propensity of NSCs. In this study, we examined the effects of controlled crosstalk between endothelial cells (ECs) and NSC neurospheres on self-renewal, differentiation, and functions of NSCs. Cultures of human fetal NSCs (hfNSCs) or human induced pluripotent stem cell (hiPSC)-derived neural progenitor cells (NPCs) in a microwell array with 500-μm well diameter facilitated cell?cell interaction and self-renewal ability, leading to increased neuronal differentiation and improved electrophysiological functions. Incorporation of ECs into size-controlled hfNSC neurospheres further promoted cell-cell interaction and self-renewal capacity. The decrease in EC density in hfNSC neurospheres effectively promoted cell?cell interaction and self-renewal. Under spontaneous differentiation condition, EC-containing hfNSC neurospheres differentiated into astrocytes rather than neuronal lineages. Therefore, we suggest the engineering of NSCs at neurosphere stage using microwell culture to control neurosphere size and that EC co-culture for vascularization may regulate behaviors, phenotypes, and functions of NSCs, leading to modulation of their therapeutic and regenerative potentials.

Keywords:Neural stem cell;Neurosphere;Microwell;Endothelial-neurosphere crosstalk;Self-renewal;Differentiation

[References]

Chan LY, Birch WR, Yim EK, Choo AB, Biomaterials, 34, 382 (2013)
Wade A, McKinney A, Phillips JJ, Biochim. Biophys. Acta, 1840, 2520 (2014)
Kazanis I, Neurobiology, 71, 1006 (2011)
Conover JC, Notti RQ, Cell Tissue Res., 331, 211 (2008)
Goldberg JS, Hirschi KK, Regener. Med., 4, 879 (2009)
Kothapalli CR, Kamm RD, Biomaterials, 34, 5995 (2013)
Jeong GS, Tissue Eng. Regener. Med., 11, 297 (2014)
Min SK, Shim HJ, Shin HS, Biotech. Bioprocess Eng., 23, 588 (2018)
Tavazoie M, Van der Veken L, Silva-Vargas V, Louissaint M, Colonna L, Zaidi B, Garcia-Verdugo JM, Doetsch F, Cell Stem Cell, 3, 279 (2008)
Hall CN, Reynell C, Gesslein B, Hamilton NB, Mishra A, Sutherland BA, O'Farrell FM, Buchan AM, Lauritzen M, Attwell D, Nature, 508(7494), 55 (2014)
Lok J, Gupta P, Guo S, Kim WJ, Whalen MJ, van Leyen K, Lo EH, Neurochem. Res., 32, 2032 (2007)
Tata M, Ruhrberg C, Fantin A, Mech. Dev., 138, 26 (2015)
Shin Y, Yang K, Han S, Park HJ, Heo YS, Cho SW, Chung S, Adv. Healthcare Mater., 3, 1457 (2014)
Kim J, Yang K, Park HJ, Cho SW, Han S, Shin Y, Chung S, Lee JH, Biotech. Bioprocess Eng., 19, 379 (2014)
Burdick JA, Vunjak-Novakovic G, Tissue Eng. A, 15, 205 (2008)
Huang F, Shen Q, Zhao J, Neural Regener. Res., 8, 313 (2013)
Dmitriev RI, Zhdanov AV, Nolan YM, Papkovsky DB, Biomaterials, 34, 9307 (2013)
Laundos TL, Silva J, Assuncao M, Quelhas P, Monteiro C, Oliveira C, Oliveira MJ, Pego AP, Amaral IF, J. Tissue Eng. Regener. Med., 11, 2227 (2017)
Dixon AR, Ramirez Y, Haengel K, Barald KF, J. Tissue Eng. Regener. Med., 12, e379 (2018)
Lee M, Yang K, Hwang YH, Byun Y, Lee DY, Cho SW, Lee H, Adv. Healthcare Mater., 4, 511 (2015)
Lee JM, Moon JY, Shaker MR, Sun W, Chung BG, Electrophoresis, 38(24), 3161 (2017)
Jeong GS, Song JH, Kang AR, Jun Y, Kim JH, Chang JY, Lee SH, Adv. Healthcare Mater., 2, 119 (2013)
Khademhosseini A, Ferreira L, Blumling J, Yeh J, Karp JM, Fukuda J, Langer R, Biomaterials, 27, 5968 (2006)
Kim HW, Yang K, Jeong WJ, Choi SJ, Lee JS, Cho AN, Chang GE, Cheong E, Cho SW, Lim YB, ACS Appl. Mater. Interfaces, 8, 26470 (2016)
Seo HI, Cho AN, Jang J, Kim DW, Cho SW, Chung BG, Nanomedicine, 11, 1861 (2015)
Kim DS, Lee DR, Kim HS, Yoo JE, Jung SJ, Lim BY, Jang J, Kang HC, You S, Hwang DY, PloS One, 7, e39715 (2012)
Yang K, Lee J, Lee JS, Kim D, Chang GE, Seo J, Cheong E, Lee T, Cho SW, ACS Appl. Mater. Interfaces, 8, 17763 (2016)
Yang K, Oh JY, Lee JS, Jin Y, Chang GE, Chae SS, Cheong E, Baik HK, Cho SW, Theranostics, 7, 4591 (2017)
Yang K, Park E, Lee JS, Kim IS, Hong K, Park KI, Cho SW, Yang HS, Macromol. Biosci., 15, 1348 (2015)
Ottone C, Krusche B, Whitby A, Clements M, Quadrato G, Pitulescu ME, Adams RH, Parrinello S, Nat. Cell Biol., 16, 1045 (2014)
Kim J, Yang K, Lee JS, Hwang YH, Park HJ, Park HJ, Lee DY, Cho SW, Macromol. Biosci., 17, 160054 (2017)
Zhang J, Woodhead GJ, Swaminathan SK, Noles SR, McQuinn ER, Pisarek AJ, Stocker AM, Mutch CA, Funatsu N, Chenn A, Dev. Cell, 18, 472 (2010)
Shen Q, Goderie SK, Jin L, Karanth N, Sun Y, Abramova N, Vincent P, Pumiglia K, Temple S, Science, 304, 1338 (2004)
Imayoshi I, Sakamoto M, Yamaguchi M, Mori K, Kageyama R, J. Neurosci., 30, 3489 (2010)
Ohtsuka T, Sakamoto M, Guillemot F, Kageyama R, J. Biol. Chem., 276, 30467 (2001)
Ohtsuka T, Ishibashi M, Gradwohl G, Nakanishi S, Guillemot F, Kageyama R, Embo J., 8, 2196 (1999)
Kageyama R, Ohtsuka T, Kobayashi T, Development, 134, 1243 (2007)
Dawson D, Volpert O, Gillis P, Crawford S, Xu HJ, Benedict W, Bouck N, Science, 285, 245 (1999)
Lee ML, Martinez-Lozada Z, Krizman EN, Robinson MB, J. Neurochem., 143, 489 (2017)
Lancaster MA, Renner M, Martin CA, Wenzel D, Bicknell LS, Hurles ME, Homfray T, Penninger JM, Jackson AP, Knoblich JA, Nature, 501(7467), 373 (2013)
Fatehullah A, Tan SH, Barker N, Nat. Cell Biol., 18, 246 (2016)
Garcez PP, Loiola EC, Da Costa RM, Higa LM, Trindade P, Delvecchio R, Nascimento JM, Brindeiro R, Tanuri A, Rehen SK, Science, 352(6287), 816 (2016)
Clevers H, Cell, 165, 1586 (2016)
Birey F, Andersen J, Makinson CD, Islam S, Wei W, Huber N, Fan HC, Metzler KRC, Panagiotakos G, Thom N, O'Rourke NA, Steinmetz LM, Bernstein JA, Hallmayer J, Huguenard JR, Pasca SP, Nature, 545(7652), 54 (2017)
Xiang Y, Tanaka Y, Patterson B, Kang YJ, Govindaiah G, Roselaar N, Cakir B, Kim KY, Lombroso AP, Hwang SM, Cell Stem Cell, 21, 383 (2017)
Jo J, Xiao Y, Sun AX, Cukuroglu E, Tran HD, Goke J, Tan ZY, Saw TY, Tan CP, Lokman H, Cell Stem Cell, 19, 248 (2016)

[1] Chan LY, Birch WR, Yim EK, Choo AB, Biomaterials, 34, 382 (2013)

[2] Wade A, McKinney A, Phillips JJ, Biochim. Biophys. Acta, 1840, 2520 (2014)

[3] Kazanis I, Neurobiology, 71, 1006 (2011)

[4] Conover JC, Notti RQ, Cell Tissue Res., 331, 211 (2008)

[5] Goldberg JS, Hirschi KK, Regener. Med., 4, 879 (2009)

[6] Kothapalli CR, Kamm RD, Biomaterials, 34, 5995 (2013)

[7] Jeong GS, Tissue Eng. Regener. Med., 11, 297 (2014)

[8] Min SK, Shim HJ, Shin HS, Biotech. Bioprocess Eng., 23, 588 (2018)

[9] Tavazoie M, Van der Veken L, Silva-Vargas V, Louissaint M, Colonna L, Zaidi B, Garcia-Verdugo JM, Doetsch F, Cell Stem Cell, 3, 279 (2008)

[10] Hall CN, Reynell C, Gesslein B, Hamilton NB, Mishra A, Sutherland BA, O'Farrell FM, Buchan AM, Lauritzen M, Attwell D, Nature, 508(7494), 55 (2014)

[11] Lok J, Gupta P, Guo S, Kim WJ, Whalen MJ, van Leyen K, Lo EH, Neurochem. Res., 32, 2032 (2007)

[12] Tata M, Ruhrberg C, Fantin A, Mech. Dev., 138, 26 (2015)

[13] Shin Y, Yang K, Han S, Park HJ, Heo YS, Cho SW, Chung S, Adv. Healthcare Mater., 3, 1457 (2014)

[14] Kim J, Yang K, Park HJ, Cho SW, Han S, Shin Y, Chung S, Lee JH, Biotech. Bioprocess Eng., 19, 379 (2014)

[15] Burdick JA, Vunjak-Novakovic G, Tissue Eng. A, 15, 205 (2008)

[16] Huang F, Shen Q, Zhao J, Neural Regener. Res., 8, 313 (2013)

[17] Dmitriev RI, Zhdanov AV, Nolan YM, Papkovsky DB, Biomaterials, 34, 9307 (2013)

[18] Laundos TL, Silva J, Assuncao M, Quelhas P, Monteiro C, Oliveira C, Oliveira MJ, Pego AP, Amaral IF, J. Tissue Eng. Regener. Med., 11, 2227 (2017)

[19] Dixon AR, Ramirez Y, Haengel K, Barald KF, J. Tissue Eng. Regener. Med., 12, e379 (2018)

[20] Lee M, Yang K, Hwang YH, Byun Y, Lee DY, Cho SW, Lee H, Adv. Healthcare Mater., 4, 511 (2015)

[21] Lee JM, Moon JY, Shaker MR, Sun W, Chung BG, Electrophoresis, 38(24), 3161 (2017)

[22] Jeong GS, Song JH, Kang AR, Jun Y, Kim JH, Chang JY, Lee SH, Adv. Healthcare Mater., 2, 119 (2013)

[23] Khademhosseini A, Ferreira L, Blumling J, Yeh J, Karp JM, Fukuda J, Langer R, Biomaterials, 27, 5968 (2006)

[24] Kim HW, Yang K, Jeong WJ, Choi SJ, Lee JS, Cho AN, Chang GE, Cheong E, Cho SW, Lim YB, ACS Appl. Mater. Interfaces, 8, 26470 (2016)

[25] Seo HI, Cho AN, Jang J, Kim DW, Cho SW, Chung BG, Nanomedicine, 11, 1861 (2015)

[26] Kim DS, Lee DR, Kim HS, Yoo JE, Jung SJ, Lim BY, Jang J, Kang HC, You S, Hwang DY, PloS One, 7, e39715 (2012)

[27] Yang K, Lee J, Lee JS, Kim D, Chang GE, Seo J, Cheong E, Lee T, Cho SW, ACS Appl. Mater. Interfaces, 8, 17763 (2016)

[28] Yang K, Oh JY, Lee JS, Jin Y, Chang GE, Chae SS, Cheong E, Baik HK, Cho SW, Theranostics, 7, 4591 (2017)

[29] Yang K, Park E, Lee JS, Kim IS, Hong K, Park KI, Cho SW, Yang HS, Macromol. Biosci., 15, 1348 (2015)

[30] Ottone C, Krusche B, Whitby A, Clements M, Quadrato G, Pitulescu ME, Adams RH, Parrinello S, Nat. Cell Biol., 16, 1045 (2014)

[31] Kim J, Yang K, Lee JS, Hwang YH, Park HJ, Park HJ, Lee DY, Cho SW, Macromol. Biosci., 17, 160054 (2017)

[32] Zhang J, Woodhead GJ, Swaminathan SK, Noles SR, McQuinn ER, Pisarek AJ, Stocker AM, Mutch CA, Funatsu N, Chenn A, Dev. Cell, 18, 472 (2010)

[33] Shen Q, Goderie SK, Jin L, Karanth N, Sun Y, Abramova N, Vincent P, Pumiglia K, Temple S, Science, 304, 1338 (2004)

[34] Imayoshi I, Sakamoto M, Yamaguchi M, Mori K, Kageyama R, J. Neurosci., 30, 3489 (2010)

[35] Ohtsuka T, Sakamoto M, Guillemot F, Kageyama R, J. Biol. Chem., 276, 30467 (2001)

[36] Ohtsuka T, Ishibashi M, Gradwohl G, Nakanishi S, Guillemot F, Kageyama R, Embo J., 8, 2196 (1999)

[37] Kageyama R, Ohtsuka T, Kobayashi T, Development, 134, 1243 (2007)

[38] Dawson D, Volpert O, Gillis P, Crawford S, Xu HJ, Benedict W, Bouck N, Science, 285, 245 (1999)

[39] Lee ML, Martinez-Lozada Z, Krizman EN, Robinson MB, J. Neurochem., 143, 489 (2017)

[40] Lancaster MA, Renner M, Martin CA, Wenzel D, Bicknell LS, Hurles ME, Homfray T, Penninger JM, Jackson AP, Knoblich JA, Nature, 501(7467), 373 (2013)

[41] Fatehullah A, Tan SH, Barker N, Nat. Cell Biol., 18, 246 (2016)

[42] Garcez PP, Loiola EC, Da Costa RM, Higa LM, Trindade P, Delvecchio R, Nascimento JM, Brindeiro R, Tanuri A, Rehen SK, Science, 352(6287), 816 (2016)

[43] Clevers H, Cell, 165, 1586 (2016)

[44] Birey F, Andersen J, Makinson CD, Islam S, Wei W, Huber N, Fan HC, Metzler KRC, Panagiotakos G, Thom N, O'Rourke NA, Steinmetz LM, Bernstein JA, Hallmayer J, Huguenard JR, Pasca SP, Nature, 545(7652), 54 (2017)

[45] Xiang Y, Tanaka Y, Patterson B, Kang YJ, Govindaiah G, Roselaar N, Cakir B, Kim KY, Lombroso AP, Hwang SM, Cell Stem Cell, 21, 383 (2017)

[46] Jo J, Xiao Y, Sun AX, Cukuroglu E, Tran HD, Goke J, Tan ZY, Saw TY, Tan CP, Lokman H, Cell Stem Cell, 19, 248 (2016)